Arginine methyltransferase inhibitors and uses thereof

ABSTRACT

Described herein are compounds of Formula (I), pharmaceutically acceptable salts thereof, and pharmaceutical compositions thereof. Compounds described herein are useful for inhibiting arginine methyltransferase activity. Methods of using the compounds for treating arginine methyltransferase-mediated disorders are also described.

RELATED APPLICATIONS

The present application is a continuation of and claims priority under35 U.S.C. § 120 to U.S. Ser. No. 15/421,699, filed Feb. 1, 2017, whichis a divisional of and claims priority under 35 U.S.C. § 121 to U.S.Ser. No. 14/775,794, filed Sep. 14, 2015, which is a national stagefiling under 35 U.S.C. § 371 of international PCT application,PCT/US2014/029710, filed Mar. 14, 2014, which claims priority under 35U.S.C. § 119(e) to U.S. provisional patent applications, U.S. Ser. No.61/781,051, filed Mar. 14, 2013, and U.S. Ser. No. 61/876,034, filedSep. 10, 2013, the entire contents of each of which is incorporatedherein by reference.

BACKGROUND OF THE INVENTION

Epigenetic regulation of gene expression is an important biologicaldeterminant of protein production and cellular differentiation and playsa significant pathogenic role in a number of human diseases.

Epigenetic regulation involves heritable modification of geneticmaterial without changing its nucleotide sequence. Typically, epigeneticregulation is mediated by selective and reversible modification (e.g.,methylation) of DNA and proteins (e.g., histones) that control theconformational transition between transcriptionally active and inactivestates of chromatin. These covalent modifications can be controlled byenzymes such as methyltransferases (e.g., arginine methyltransferases),many of which are associated with specific genetic alterations that cancause human disease.

Disease-associated chromatin-modifying enzymes (e.g., argininemethyltransferases) play a role in diseases such as proliferativedisorders, autoimmune disorders, muscular disorders, vascular disorders,metabolic disorders, and neurological disorders. Thus, there is a needfor the development of small molecules that are capable of inhibitingthe activity of arginine methyltransferases.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS

Arginine methyltransferases are attractive targets for modulation giventheir role in the regulation of diverse biological processes. It has nowbeen found that compounds described herein, and pharmaceuticallyacceptable salts and compositions thereof, are effective as inhibitorsof arginine methyltransferases. Such compounds have the general Formula(I):

or a pharmaceutically acceptable salt thereof, wherein X, Y, Z, L₁,R^(W), R³, and R^(x) are as defined herein.

In some embodiments, pharmaceutical compositions are provided whichcomprise a compound described herein (e.g., a compound of Formula (I)),or a pharmaceutically acceptable salt thereof, and optionally apharmaceutically acceptable excipient.

In certain embodiments, compounds described herein inhibit activity ofan arginine methyltransferase (RMT) (e.g., PRMT1, PRMT3, CARM1, PRMT6,and/or PRMT8). In certain embodiments, methods of inhibiting an argininemethyltransferase are provided which comprise contacting the argininemethyltransferase with an effective amount of a compound of Formula (I),or a pharmaceutically acceptable salt thereof. The RMT may be purifiedor crude, and may be present in a cell, tissue, or a subject. Thus, suchmethods encompass inhibition of RMT activity both in vitro and in vivo.In certain embodiments, the RMT is wild-type. In certain embodiments,the RMT is overexpressed. In certain embodiments, the RMT is a mutant.In certain embodiments, the RMT is in a cell. In some embodiments, theRMT is expressed at normal levels in a subject, but the subject wouldbenefit from RMT inhibition (e.g., because the subject has one or moremutations in an RMT substrate that causes an increase in methylation ofthe substrate with normal levels of RMT). In some embodiments, the RMTis in a subject known or identified as having abnormal RMT activity(e.g., overexpression).

In certain embodiments, methods of modulating gene expression in a cellare provided which comprise contacting a cell with an effective amountof a compound of Formula (I), or a pharmaceutically acceptable saltthereof, or a pharmaceutical composition thereof. In certainembodiments, the cell in culture in vitro. In certain embodiments, cellis in an animal, e.g., a human.

In certain embodiments, methods of modulating transcription in a cellare provided which comprise contacting a cell with an effective amountof a compound of Formula (I), or a pharmaceutically acceptable saltthereof, or a pharmaceutical composition thereof. In certainembodiments, the cell in culture in vitro. In certain embodiments, thecell is in an animal, e.g., a human.

In some embodiments, methods of treating an RMT-mediated disorder (e.g.,a PRMT1-, PRMT3-, CARM1-, PRMT6-, or PRMT8-mediated disorder) areprovided which comprise administering to a subject suffering from anRMT-mediated disorder an effective amount of a compound described herein(e.g., a compound of Formula (I)), or a pharmaceutically acceptable saltthereof, or a pharmaceutical composition thereof. In certainembodiments, the RMT-mediated disorder is a proliferative disorder. Incertain embodiments, compounds described herein are useful for treatingcancer. In certain embodiments, compounds described herein are usefulfor treating breast cancer, prostate cancer, lung cancer, colon cancer,bladder cancer, or leukemia. In certain embodiments, the RMT-mediateddisorder is a muscular disorder. In certain embodiments, theRMT-mediated disorder is an autoimmune disorder. In certain embodiments,the RMT-mediated disorder is a neurological disorder. In certainembodiments, the RMT-mediated disorder is a vascular disorder. Incertain embodiments, the RMT-mediated disorder is a metabolic disorder.

Compounds described herein are also useful for the study of argininemethyltransferases in biological and pathological phenomena, the studyof intracellular signal transduction pathways mediated by argininemethyltransferases, and the comparative evaluation of new RMTinhibitors.

This application refers to various issued patent, published patentapplications, journal articles, and other publications, all of which areincorporated herein by reference.

Definitions of specific functional groups and chemical terms aredescribed in more detail below. The chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75^(th) Ed., inside cover, andspecific functional groups are generally defined as described therein.Additionally, general principles of organic chemistry, as well asspecific functional moieties and reactivity, are described in ThomasSorrell, Organic Chemistry, University Science Books, Sausalito, 1999;Smith and March, March's Advanced Organic Chemistry, 5^(th) Edition,John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive OrganicTransformations, VCH Publishers, Inc., New York, 1989; and Carruthers,Some Modern Methods of Organic Synthesis, 3^(rd) Edition, CambridgeUniversity Press, Cambridge, 1987.

Compounds described herein can comprise one or more asymmetric centers,and thus can exist in various isomeric forms, e.g., enantiomers and/ordiastereomers. For example, the compounds described herein can be in theform of an individual enantiomer, diastereomer or geometric isomer, orcan be in the form of a mixture of stereoisomers, including racemicmixtures and mixtures enriched in one or more stereoisomer. Isomers canbe isolated from mixtures by methods known to those skilled in the art,including chiral high pressure liquid chromatography (HPLC) and theformation and crystallization of chiral salts; or preferred isomers canbe prepared by asymmetric syntheses. See, for example, Jacques et al.,Enantiomers, Racemates and Resolutions (Wiley Interscience, New York,1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistryof Carbon Compounds (McGraw-Hill, N Y, 1962); and Wilen, Tables ofResolving Agents and Optical Resolutions p. 268 (E. L. Eliel, Ed., Univ.of Notre Dame Press, Notre Dame, Ind. 1972). The present disclosureadditionally encompasses compounds described herein as individualisomers substantially free of other isomers, and alternatively, asmixtures of various isomers.

It is to be understood that the compounds of the present invention maybe depicted as different tautomers. It should also be understood thatwhen compounds have tautomeric forms, all tautomeric forms are intendedto be included in the scope of the present invention, and the naming ofany compound described herein does not exclude any tautomer form.

Unless otherwise stated, structures depicted herein are also meant toinclude compounds that differ only in the presence of one or moreisotopically enriched atoms. For example, compounds having the presentstructures except for the replacement of hydrogen by deuterium ortritium, replacement of ¹⁹F with ¹⁸F, or the replacement of a carbon bya ¹³C- or ¹⁴C-enriched carbon are within the scope of the disclosure.Such compounds are useful, for example, as analytical tools or probes inbiological assays.

When a range of values is listed, it is intended to encompass each valueand sub-range within the range. For example “C₁₋₆ alkyl” is intended toencompass, C₁, C₂, C₃, C₄, C₅, C₆, C₁₋₆, C₁₋₅, C₁₋₄, C₁₋₃, C₁₋₂, C₂₋₆,C₂₋₅, C₂₋₄, C₂₋₃, C₃₋₆, C₃₋₅, C₃₋₄, C₄₋₆, C₄₋₅, and C₅₋₆ alkyl.

“Radical” refers to a point of attachment on a particular group. Radicalincludes divalent radicals of a particular group.

“Alkyl” refers to a radical of a straight-chain or branched saturatedhydrocarbon group having from 1 to 20 carbon atoms (“C₁₋₂₀ alkyl”). Insome embodiments, an alkyl group has 1 to 10 carbon atoms (“C₁₋₁₀alkyl”). In some embodiments, an alkyl group has 1 to 9 carbon atoms(“C₁₋₉ alkyl”). In some embodiments, an alkyl group has 1 to 8 carbonatoms (“C₁₋₈ alkyl”). In some embodiments, an alkyl group has 1 to 7carbon atoms (“C₁₋₇ alkyl”). In some embodiments, an alkyl group has 1to 6 carbon atoms (“C₁₋₆ alkyl”). In some embodiments, an alkyl grouphas 1 to 5 carbon atoms (“C₁₋₅ alkyl”). In some embodiments, an alkylgroup has 1 to 4 carbon atoms (“C₁₋₄ alkyl”). In some embodiments, analkyl group has 1 to 3 carbon atoms (“C₁₋₃ alkyl”). In some embodiments,an alkyl group has 1 to 2 carbon atoms (“C₁₋₂ alkyl”). In someembodiments, an alkyl group has 1 carbon atom (“C₁ alkyl”). In someembodiments, an alkyl group has 2 to 6 carbon atoms (“C₂₋₆ alkyl”).Examples of C₁₋₆ alkyl groups include methyl (C₁), ethyl (C₂), n-propyl(C₃), isopropyl (C₃), n-butyl (C₄), tert-butyl (C₄), sec-butyl (C₄),iso-butyl (C₄), n-pentyl (C₅), 3-pentanyl (C₅), amyl (C₅), neopentyl(C₅), 3-methyl-2-butanyl (C₅), tertiary amyl (C₅), and n-hexyl (C₆).Additional examples of alkyl groups include n-heptyl (C₇), n-octyl (C₈)and the like. In certain embodiments, each instance of an alkyl group isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted alkyl”) or substituted (a “substituted alkyl”) with oneor more substituents. In certain embodiments, the alkyl group isunsubstituted C₁₋₁₀ alkyl (e.g., —CH₃). In certain embodiments, thealkyl group is substituted C₁₋₁₀ alkyl.

In some embodiments, an alkyl group is substituted with one or morehalogens. “Perhaloalkyl” is a substituted alkyl group as defined hereinwherein all of the hydrogen atoms are independently replaced by ahalogen, e.g., fluoro, bromo, chloro, or iodo. In some embodiments, thealkyl moiety has 1 to 8 carbon atoms (“C₁₋₈ perhaloalkyl”). In someembodiments, the alkyl moiety has 1 to 6 carbon atoms (“C₁₋₆perhaloalkyl”). In some embodiments, the alkyl moiety has 1 to 4 carbonatoms (“C₁₋₄ perhaloalkyl”). In some embodiments, the alkyl moiety has 1to 3 carbon atoms (“C₁₋₃ perhaloalkyl”). In some embodiments, the alkylmoiety has 1 to 2 carbon atoms (“C₁₋₂ perhaloalkyl”). In someembodiments, all of the hydrogen atoms are replaced with fluoro. In someembodiments, all of the hydrogen atoms are replaced with chloro.Examples of perhaloalkyl groups include —CF₃, —CF₂CF₃, —CF₂CF₂CF₃,—CCl₃, —CFCl₂, —CF₂Cl, and the like.

“Alkenyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms and one or morecarbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds), andoptionally one or more triple bonds (e.g., 1, 2, 3, or 4 triple bonds)(“C₂₋₂₀ alkenyl”). In certain embodiments, alkenyl does not comprisetriple bonds. In some embodiments, an alkenyl group has 2 to 10 carbonatoms (“C₂₋₁₀ alkenyl”). In some embodiments, an alkenyl group has 2 to9 carbon atoms (“C₂₋₉ alkenyl”). In some embodiments, an alkenyl grouphas 2 to 8 carbon atoms (“C₂₋₈ alkenyl”). In some embodiments, analkenyl group has 2 to 7 carbon atoms (“C₂₋₇ alkenyl”). In someembodiments, an alkenyl group has 2 to 6 carbon atoms (“C₂₋₆ alkenyl”).In some embodiments, an alkenyl group has 2 to 5 carbon atoms (“C₂₋₅alkenyl”). In some embodiments, an alkenyl group has 2 to 4 carbon atoms(“C₂₋₄ alkenyl”). In some embodiments, an alkenyl group has 2 to 3carbon atoms (“C₂₋₃ alkenyl”). In some embodiments, an alkenyl group has2 carbon atoms (“C₂ alkenyl”). The one or more carbon-carbon doublebonds can be internal (such as in 2-butenyl) or terminal (such as in1-butenyl). Examples of C₂₋₄ alkenyl groups include ethenyl (C₂),1-propenyl (C₃), 2-propenyl (C₃), 1-butenyl (C₄), 2-butenyl (C₄),butadienyl (C₄), and the like. Examples of C₂₋₆ alkenyl groups includethe aforementioned C₂₋₄ alkenyl groups as well as pentenyl (C₅),pentadienyl (C₅), hexenyl (C₆), and the like. Additional examples ofalkenyl include heptenyl (C₇), octenyl (C₈), octatrienyl (C₈), and thelike. In certain embodiments, each instance of an alkenyl group isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted alkenyl”) or substituted (a “substituted alkenyl”) withone or more substituents. In certain embodiments, the alkenyl group isunsubstituted C₂₋₁₀ alkenyl. In certain embodiments, the alkenyl groupis substituted C₂₋₁₀ alkenyl.

“Alkynyl” refers to a radical of a straight-chain or branchedhydrocarbon group having from 2 to 20 carbon atoms and one or morecarbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds), andoptionally one or more double bonds (e.g., 1, 2, 3, or 4 double bonds)(“C₂₋₂₀ alkynyl”). In certain embodiments, alkynyl does not comprisedouble bonds. In some embodiments, an alkynyl group has 2 to 10 carbonatoms (“C₂₋₁₀ alkynyl”). In some embodiments, an alkynyl group has 2 to9 carbon atoms (“C₂₋₉ alkynyl”). In some embodiments, an alkynyl grouphas 2 to 8 carbon atoms (“C₂₋₈ alkynyl”). In some embodiments, analkynyl group has 2 to 7 carbon atoms (“C₂₋₇ alkynyl”). In someembodiments, an alkynyl group has 2 to 6 carbon atoms (“C₂₋₆ alkynyl”).In some embodiments, an alkynyl group has 2 to 5 carbon atoms (“C₂₋₅alkynyl”). In some embodiments, an alkynyl group has 2 to 4 carbon atoms(“C₂₋₄ alkynyl”). In some embodiments, an alkynyl group has 2 to 3carbon atoms (“C₂₋₃ alkynyl”). In some embodiments, an alkynyl group has2 carbon atoms (“C₂ alkynyl”). The one or more carbon-carbon triplebonds can be internal (such as in 2-butynyl) or terminal (such as in1-butynyl). Examples of C₂₋₄ alkynyl groups include, without limitation,ethynyl (C₂), 1-propynyl (C₃), 2-propynyl (C₃), 1-butynyl (C₄),2-butynyl (C₄), and the like. Examples of C₂₋₆ alkenyl groups includethe aforementioned C₂₋₄ alkynyl groups as well as pentynyl (C₅), hexynyl(C₆), and the like. Additional examples of alkynyl include heptynyl(C₇), octynyl (C₈), and the like. In certain embodiments, each instanceof an alkynyl group is independently optionally substituted, e.g.,unsubstituted (an “unsubstituted alkynyl”) or substituted (a“substituted alkynyl”) with one or more substituents. In certainembodiments, the alkynyl group is unsubstituted C₂₋₁₀ alkynyl. Incertain embodiments, the alkynyl group is substituted C₂₋₁₀ alkynyl.

“Fused” or “ortho-fused” are used interchangeably herein, and refer totwo rings that have two atoms and one bond in common, e.g.,

“Bridged” refers to a ring system containing (1) a bridgehead atom orgroup of atoms which connect two or more non-adjacent positions of thesame ring; or (2) a bridgehead atom or group of atoms which connect twoor more positions of different rings of a ring system and does notthereby form an ortho-fused ring, e.g.,

“Spiro” or “Spiro-fused” refers to a group of atoms which connect to thesame atom of a carbocyclic or heterocyclic ring system (geminalattachment), thereby forming a ring, e.g.,

Spiro-fusion at a bridgehead atom is also contemplated.

“Carbocyclyl” or “carbocyclic” refers to a radical of a non-aromaticcyclic hydrocarbon group having from 3 to 14 ring carbon atoms (“C₃₋₁₄carbocyclyl”) and zero heteroatoms in the non-aromatic ring system. Incertain embodiments, a carbocyclyl group refers to a radical of anon-aromatic cyclic hydrocarbon group having from 3 to 10 ring carbonatoms (“C₃₋₁₀ carbocyclyl”) and zero heteroatoms in the non-aromaticring system. In some embodiments, a carbocyclyl group has 3 to 8 ringcarbon atoms (“C₃₋₈ carbocyclyl”). In some embodiments, a carbocyclylgroup has 3 to 6 ring carbon atoms (“C₃₋₆ carbocyclyl”). In someembodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C₃₋₆carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 10 ringcarbon atoms (“C₅₋₁₀ carbocyclyl”). Exemplary C₃₋₆ carbocyclyl groupsinclude, without limitation, cyclopropyl (C₃), cyclopropenyl (C₃),cyclobutyl (C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl(C₅), cyclohexyl (C₆), cyclohexenyl (C₆), cyclohexadienyl (C₆), and thelike. Exemplary C₃₋₈ carbocyclyl groups include, without limitation, theaforementioned C₃₋₆ carbocyclyl groups as well as cycloheptyl (C₇),cycloheptenyl (C₇), cycloheptadienyl (C₇), cycloheptatrienyl (C₇),cyclooctyl (C₈), cyclooctenyl (C₈), bicyclo[2.2.1]heptanyl (C₇),bicyclo[2.2.2]octanyl (C₈), and the like. Exemplary C₃₋₁₀ carbocyclylgroups include, without limitation, the aforementioned C₃₋₈ carbocyclylgroups as well as cyclononyl (C₉), cyclononenyl (C₉), cyclodecyl (C₁₀),cyclodecenyl (C₁₀), octahydro-1H-indenyl (C₉), decahydronaphthalenyl(C₁₀), spiro[4.5]decanyl (C₁₀), and the like. As the foregoing examplesillustrate, in certain embodiments, the carbocyclyl group is eithermonocyclic (“monocyclic carbocyclyl”) or is a fused, bridged orspiro-fused ring system such as a bicyclic system (“bicycliccarbocyclyl”) and can be saturated or can be partially unsaturated.“Carbocyclyl” also includes ring systems wherein the carbocyclyl ring,as defined above, is fused with one or more aryl or heteroaryl groupswherein the point of attachment is on the carbocyclyl ring, and in suchinstances, the number of carbons continue to designate the number ofcarbons in the carbocyclic ring system. In certain embodiments, eachinstance of a carbocyclyl group is independently optionally substituted,e.g., unsubstituted (an “unsubstituted carbocyclyl”) or substituted (a“substituted carbocyclyl”) with one or more substituents. In certainembodiments, the carbocyclyl group is unsubstituted C₃₋₁₀ carbocyclyl.In certain embodiments, the carbocyclyl group is a substituted C₃₋₁₀carbocyclyl.

In some embodiments, “carbocyclyl” is a monocyclic, saturatedcarbocyclyl group having from 3 to 14 ring carbon atoms (“C₃₋₁₄cycloalkyl”). In some embodiments, “carbocyclyl” is a monocyclic,saturated carbocyclyl group having from 3 to 10 ring carbon atoms(“C₃₋₁₀ cycloalkyl”). In some embodiments, a cycloalkyl group has 3 to 8ring carbon atoms (“C₃₋₈ cycloalkyl”). In some embodiments, a cycloalkylgroup has 3 to 6 ring carbon atoms (“C₃₋₆ cycloalkyl”). In someembodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C₅₋₆cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 10 ringcarbon atoms (“C₅₋₁₀ cycloalkyl”). Examples of C₅₋₆ cycloalkyl groupsinclude cyclopentyl (C₅) and cyclohexyl (C₅). Examples of C₃₋₆cycloalkyl groups include the aforementioned C₅₋₆ cycloalkyl groups aswell as cyclopropyl (C₃) and cyclobutyl (C₄). Examples of C₃₋₈cycloalkyl groups include the aforementioned C₃₋₆ cycloalkyl groups aswell as cycloheptyl (C₇) and cyclooctyl (C₈). In certain embodiments,each instance of a cycloalkyl group is independently unsubstituted (an“unsubstituted cycloalkyl”) or substituted (a “substituted cycloalkyl”)with one or more substituents. In certain embodiments, the cycloalkylgroup is unsubstituted C₃₋₁₀ cycloalkyl. In certain embodiments, thecycloalkyl group is substituted C₃₋₁₀ cycloalkyl.

“Heterocyclyl” or “heterocyclic” refers to a radical of a 3-to14-membered non-aromatic ring system having ring carbon atoms and 1 to 4ring heteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“3-14 membered heterocyclyl”). In certainembodiments, heterocyclyl or heterocyclic refers to a radical of a 3-10membered non-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“3-10 membered heterocyclyl”). Inheterocyclyl groups that contain one or more nitrogen atoms, the pointof attachment can be a carbon or nitrogen atom, as valency permits. Aheterocyclyl group can either be monocyclic (“monocyclic heterocyclyl”)or a fused, bridged or spiro-fused ring system such as a bicyclic system(“bicyclic heterocyclyl”), and can be saturated or can be partiallyunsaturated. Heterocyclyl bicyclic ring systems can include one or moreheteroatoms in one or both rings. “Heterocyclyl” also includes ringsystems wherein the heterocyclyl ring, as defined above, is fused withone or more carbocyclyl groups wherein the point of attachment is eitheron the carbocyclyl or heterocyclyl ring, or ring systems wherein theheterocyclyl ring, as defined above, is fused with one or more aryl orheteroaryl groups, wherein the point of attachment is on theheterocyclyl ring, and in such instances, the number of ring memberscontinue to designate the number of ring members in the heterocyclylring system. In certain embodiments, each instance of heterocyclyl isindependently optionally substituted, e.g., unsubstituted (an“unsubstituted heterocyclyl”) or substituted (a “substitutedheterocyclyl”) with one or more substituents. In certain embodiments,the heterocyclyl group is unsubstituted 3-10 membered heterocyclyl. Incertain embodiments, the heterocyclyl group is substituted 3-10 memberedheterocyclyl.

In some embodiments, a heterocyclyl group is a 5-10 memberednon-aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms, wherein each heteroatom is independently selected fromnitrogen, oxygen, and sulfur (“5-10 membered heterocyclyl”). In someembodiments, a heterocyclyl group is a 5-8 membered non-aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms, wherein eachheteroatom is independently selected from nitrogen, oxygen, and sulfur(“5-8 membered heterocyclyl”). In some embodiments, a heterocyclyl groupis a 5-6 membered non-aromatic ring system having ring carbon atoms and1-4 ring heteroatoms, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heterocyclyl”). In someembodiments, the 5-6 membered heterocyclyl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heterocyclyl has one ring heteroatomselected from nitrogen, oxygen, and sulfur.

Exemplary 3-membered heterocyclyl groups containing one heteroatominclude, without limitation, azirdinyl, oxiranyl, and thiiranyl.Exemplary 4-membered heterocyclyl groups containing one heteroatominclude, without limitation, azetidinyl, oxetanyl, and thietanyl.Exemplary 5-membered heterocyclyl groups containing one heteroatominclude, without limitation, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl,and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclyl groupscontaining two heteroatoms include, without limitation, dioxolanyl,oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-memberedheterocyclyl groups containing three heteroatoms include, withoutlimitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary6-membered heterocyclyl groups containing one heteroatom include,without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl,and thianyl. Exemplary 6-membered heterocyclyl groups containing twoheteroatoms include, without limitation, piperazinyl, morpholinyl,dithianyl, and dioxanyl. Exemplary 6-membered heterocyclyl groupscontaining three heteroatoms include, without limitation, triazinanyl.Exemplary 7-membered heterocyclyl groups containing one heteroatominclude, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary8-membered heterocyclyl groups containing one heteroatom include,without limitation, azocanyl, oxecanyl, and thiocanyl. Exemplary5-membered heterocyclyl groups fused to a C₆ aryl ring (also referred toherein as a 5,6-bicyclic heterocyclic ring) include, without limitation,indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl,benzoxazolinonyl, and the like. Exemplary 6-membered heterocyclyl groupsfused to an aryl ring (also referred to herein as a 6,6-bicyclicheterocyclic ring) include, without limitation, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and the like.

“Aryl” refers to a radical of a monocyclic or polycyclic (e.g., bicyclicor tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 πelectrons shared in a cyclic array) having 6-14 ring carbon atoms andzero heteroatoms provided in the aromatic ring system (“C₆₋₁₄ aryl”). Insome embodiments, an aryl group has six ring carbon atoms (“C₆ aryl”;e.g., phenyl). In some embodiments, an aryl group has ten ring carbonatoms (“C₁₀ aryl”; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). Insome embodiments, an aryl group has fourteen ring carbon atoms (“C₁₄aryl”; e.g., anthracyl). “Aryl” also includes ring systems wherein thearyl ring, as defined above, is fused with one or more carbocyclyl orheterocyclyl groups wherein the radical or point of attachment is on thearyl ring, and in such instances, the number of carbon atoms continue todesignate the number of carbon atoms in the aryl ring system. In certainembodiments, each instance of an aryl group is independently optionallysubstituted, e.g., unsubstituted (an “unsubstituted aryl”) orsubstituted (a “substituted aryl”) with one or more substituents. Incertain embodiments, the aryl group is unsubstituted C₆₋₁₄ aryl. Incertain embodiments, the aryl group is substituted C₆₋₁₄ aryl.

“Heteroaryl” refers to a radical of a 5-14 membered monocyclic orpolycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system(e.g., having 6 or 10 π electrons shared in a cyclic array) having ringcarbon atoms and 1-4 ring heteroatoms provided in the aromatic ringsystem, wherein each heteroatom is independently selected from nitrogen,oxygen, and sulfur (“5-14 membered heteroaryl”). In certain embodiments,heteroaryl refers to a radical of a 5-10 membered monocyclic or bicyclic4n+2 aromatic ring system having ring carbon atoms and 1-4 ringheteroatoms provided in the aromatic ring system, wherein eachheteroatom is independently selected from nitrogen, oxygen and sulfur(“5-10 membered heteroaryl”). In heteroaryl groups that contain one ormore nitrogen atoms, the point of attachment can be a carbon or nitrogenatom, as valency permits. Heteroaryl bicyclic ring systems can includeone or more heteroatoms in one or both rings. “Heteroaryl” includes ringsystems wherein the heteroaryl ring, as defined above, is fused with oneor more carbocyclyl or heterocyclyl groups wherein the point ofattachment is on the heteroaryl ring, and in such instances, the numberof ring members continue to designate the number of ring members in theheteroaryl ring system. “Heteroaryl” also includes ring systems whereinthe heteroaryl ring, as defined above, is fused with one or more arylgroups wherein the point of attachment is either on the aryl orheteroaryl ring, and in such instances, the number of ring membersdesignates the number of ring members in the fused (aryl/heteroaryl)ring system. Bicyclic heteroaryl groups wherein one ring does notcontain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and thelike) the point of attachment can be on either ring, e.g., either thering bearing a heteroatom (e.g., 2-indolyl) or the ring that does notcontain a heteroatom (e.g., 5-indolyl).

In some embodiments, a heteroaryl group is a 5-14 membered aromatic ringsystem having ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-14 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-10 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-10 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-8 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-8 membered heteroaryl”). In someembodiments, a heteroaryl group is a 5-6 membered aromatic ring systemhaving ring carbon atoms and 1-4 ring heteroatoms provided in thearomatic ring system, wherein each heteroatom is independently selectedfrom nitrogen, oxygen, and sulfur (“5-6 membered heteroaryl”). In someembodiments, the 5-6 membered heteroaryl has 1-3 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1-2 ring heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selectedfrom nitrogen, oxygen, and sulfur. In certain embodiments, each instanceof a heteroaryl group is independently optionally substituted, e.g.,unsubstituted (“unsubstituted heteroaryl”) or substituted (“substitutedheteroaryl”) with one or more substituents. In certain embodiments, theheteroaryl group is unsubstituted 5-14 membered heteroaryl. In certainembodiments, the heteroaryl group is substituted 5-14 memberedheteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatominclude, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary5-membered heteroaryl groups containing two heteroatoms include, withoutlimitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, andisothiazolyl. Exemplary 5-membered heteroaryl groups containing threeheteroatoms include, without limitation, triazolyl, oxadiazolyl, andthiadiazolyl. Exemplary 5-membered heteroaryl groups containing fourheteroatoms include, without limitation, tetrazolyl. Exemplary6-membered heteroaryl groups containing one heteroatom include, withoutlimitation, pyridinyl. Exemplary 6-membered heteroaryl groups containingtwo heteroatoms include, without limitation, pyridazinyl, pyrimidinyl,and pyrazinyl. Exemplary 6-membered heteroaryl groups containing threeor four heteroatoms include, without limitation, triazinyl andtetrazinyl, respectively. Exemplary 7-membered heteroaryl groupscontaining one heteroatom include, without limitation, azepinyl,oxepinyl, and thiepinyl. Exemplary 6,6-bicyclic heteroaryl groupsinclude, without limitation, naphthyridinyl, pteridinyl, quinolinyl,isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl.Exemplary 5,6-bicyclic heteroaryl groups include, without limitation,any one of the following formulae:

In any of the monocyclic or bicyclic heteroaryl groups, the point ofattachment can be any carbon or nitrogen atom, as valency permits.

“Partially unsaturated” refers to a group that includes at least onedouble or triple bond. The term “partially unsaturated” is intended toencompass rings having multiple sites of unsaturation, but is notintended to include aromatic groups (e.g., aryl or heteroaryl groups) asherein defined. Likewise, “saturated” refers to a group that does notcontain a double or triple bond, i.e., contains all single bonds.

In some embodiments, alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl,aryl, and heteroaryl groups, as defined herein, are optionallysubstituted (e.g., “substituted” or “unsubstituted” alkyl, “substituted”or “unsubstituted” alkenyl, “substituted” or “unsubstituted” alkynyl,“substituted” or “unsubstituted” carbocyclyl, “substituted” or“unsubstituted” heterocyclyl, “substituted” or “unsubstituted” aryl or“substituted” or “unsubstituted” heteroaryl group). In general, the term“substituted”, whether preceded by the term “optionally” or not, meansthat at least one hydrogen present on a group (e.g., a carbon ornitrogen atom) is replaced with a permissible substituent, e.g., asubstituent which upon substitution results in a stable compound, e.g.,a compound which does not spontaneously undergo transformation such asby rearrangement, cyclization, elimination, or other reaction. Unlessotherwise indicated, a “substituted” group has a substituent at one ormore substitutable positions of the group, and when more than oneposition in any given structure is substituted, the substituent iseither the same or different at each position. The term “substituted” iscontemplated to include substitution with all permissible substituentsof organic compounds, including any of the substituents described hereinthat results in the formation of a stable compound. The presentdisclosure contemplates any and all such combinations in order to arriveat a stable compound. For purposes of this disclosure, heteroatoms suchas nitrogen may have hydrogen substituents and/or any suitablesubstituent as described herein which satisfy the valencies of theheteroatoms and results in the formation of a stable moiety.

Exemplary carbon atom substituents include, but are not limited to,halogen, —CN, —NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(aa), —ON(R^(bb))₂,—N(R^(bb))₂, —N(R^(bb))₃ ⁺X⁻, —N(OR^(cc))R^(bb), —SH, —SR^(aa),—SSR^(cc), —C(═O)R^(aa), —CO₂H, —CHO, —C(OR^(cc))₂, —CO₂R^(aa),—OC(═O)R^(aa), —OCO₂R^(aa), —C(═O)N(R^(bb))₂, —OC(═O)N(R^(bb))₂,—NR^(bb)C(═O)R^(aa), —NR^(bb)CO₂R^(aa), NR^(bb)C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —OC(═NR^(bb))R^(aa),—OC(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂, —OC(═NR^(bb))N(R^(bb))₂,—NR^(bb)C(═NR^(bb))N(R^(bb))₂, —C(═O)NR^(bb)SO₂R^(aa),—NR^(bb)SO₂R^(aa), —SO₂N(R^(bb))₂, —SO₂R^(aa), —SO₂OR^(aa), —OSO₂R^(aa),—S(═O)R^(aa), —OS(═O)R^(aa), —Si(R^(aa))₃,—OSi(R^(aa))₃—C(═S)N(R^(bb))₂, —C(═O)SR^(aa), —C(═S)SR^(aa),—SC(═S)SR^(aa), —SC(═O)SR^(aa), —OC(═O)SR^(aa), —SC(═O)OR^(aa),—SC(═O)R^(aa), —P(═O)₂R^(aa), —OP(═O)₂R^(aa), —P(═O)(R^(aa))₂,—OP(═O)(R^(aa))₂, —OP(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂,—OP(═O)₂N(R^(bb))₂, —P(═O)(NR^(bb))₂, —OP(═O)(NR^(bb))₂,—NR^(bb)P(═O)(OR^(cc))₂, —NR^(bb)P(═O)(NR^(bb))₂, —P(R^(cc))₂,—P(R^(cc))₃, —OP(R^(cc))₂, —OP(R^(cc))₃, —B(R^(aa))₂, —B(OR^(cc))₂,—BR^(aa)(OR^(cc)), C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

or two geminal hydrogens on a carbon atom are replaced with the group═O, ═S, ═NN(R^(bb))₂, ═NNR^(bb)C(═O)R^(aa), ═NNR^(bb)C(═O)OR^(aa),═NNR^(bb)S(═O)₂R^(aa), ═NR^(bb), or ═NOR^(cc);

each instance of R^(aa) is, independently, selected from C₁₋₁₀ alkyl,C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀ carbocyclyl,3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 membered heteroaryl, ortwo R^(aa) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(bb) is, independently, selected from hydrogen, —OH,—OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂,—SO₂R^(cc), —SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc),—C(═S)SR^(cc), —P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂,—P(═O)(NR^(cc))₂, C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀alkynyl, C₃₋₁₀ carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and5-14 membered heteroaryl, or two R^(bb) groups are joined to form a 3-14membered heterocyclyl or 5-14 membered heteroaryl ring, wherein eachalkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroarylis independently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(cc) is, independently, selected from hydrogen, C₁₋₁₀alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups are joined to form a 3-14 memberedheterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl,alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl isindependently substituted with 0, 1, 2, 3, 4, or 5 R^(dd) groups;

each instance of R^(dd) is, independently, selected from halogen, —CN,—NO₂, —N₃, —SO₂H, —SO₃H, —OH, —OR^(ee), —ON(R^(ff))₂, —N(R^(ff))₂,—N(R^(ff))₃ ⁺X⁻, —N(OR^(ee))R^(ff), —SH, —SR^(ee), —SSR^(ee),—C(═O)R^(ee), —CO₂H, —CO₂R^(ee), —OC(═O)R^(ee), —OCO₂R^(ee),—C(═O)N(R^(ff))₂, —OC(═O)N(R^(ff))₂, —NR^(ff)C(═O)R^(ee),—NR^(ff)CO₂R^(ee), —NRC^(ff)(═O)N(R^(ff))₂, —C(═NR^(ff))OR^(ee),—OC(═NR^(ff))R^(ee), —OC(═NR^(ff))OR^(ee), —C(═NR^(ff))N(R^(ff))₂,—OC(═NR^(ff))N(R^(ff))₂, —NR^(ff)C(═NR^(ff))N(R^(ff))₂,—NR^(ff)SO₂R^(ee), —SO₂N(R)₂, —SO₂R^(ee), —SO₂OR^(ee), —OSO₂R^(ee),—S(═O)R^(ee), —Si(R^(ee))₃, —OSi(R^(ee))₃, —C(═S)N(R^(ff))₂,—C(═O)SR^(ee), —C(═S)SR^(ee), —SC(═S)SR^(ee), —P(═O)₂R^(ee),—P(═O)(R^(ee))₂, —OP(═O)(R^(ee))₂, —OP(═O)(OR^(ee))₂, C₁₋₆ alkyl, C₁₋₆perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, 3-10membered heterocyclyl, C₆₋₁₀ aryl, 5-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups, or two geminal R^(dd) substituents can be joined to form ═O or═S;

each instance of R^(ee) is, independently, selected from C₁₋₆ alkyl,C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl, C₆₋₁₀aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, whereineach alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, andheteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 R^(gg)groups;

each instance of R^(ff) is, independently, selected from hydrogen, C₁₋₆alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀ carbocyclyl,3-10 membered heterocyclyl, C₆₋₁₀ aryl and 5-10 membered heteroaryl, ortwo R^(ff) groups are joined to form a 3-14 membered heterocyclyl or5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl,carbocyclyl, heterocyclyl, aryl, and heteroaryl is independentlysubstituted with 0, 1, 2, 3, 4, or 5 R^(gg) groups; and

each instance of R^(gg) is, independently, halogen, —CN, —NO₂, —N₃,—SO₂H, —SO₃H, —OH, —OC₁₋₆ alkyl, —ON(C₁₋₆ alkyl)₂, —N(C₁₋₆ alkyl)₂,—N(C₁₋₆ alkyl)₃ ⁺X⁻, —NH(C₁₋₆ alkyl)₂ ⁺X⁻, —NH₂(C₁₋₆ alkyl)⁺X⁻, —NH₃⁺X⁻, —N(OC₁₋₆ alkyl)(C₁₋₆ alkyl), —N(OH)(C₁₋₆ alkyl), —NH(OH), —SH,—SC₁₋₆ alkyl, —SS(C₁₋₆ alkyl), —C(═O)(C₁₋₆ alkyl), —CO₂H, —CO₂(C₁₋₆alkyl), —OC(═O)(C₁₋₆ alkyl), —OCO₂(C₁₋₆ alkyl), —C(═O)NH₂, —C(═O)N(C₁₋₆alkyl)₂, —OC(═O)NH(C₁₋₆ alkyl), —NHC(═O)(C₁₋₆ alkyl), —N(C₁₋₆alkyl)C(═O)(C₁₋₆ alkyl), —NHCO₂(C₁₋₆ alkyl), —NHC(═O)N(C₁₋₆ alkyl)₂,—NHC(═O)NH(C₁₋₆ alkyl), —NHC(═O)NH₂, —C(═NH)O(C₁₋₆ alkyl), —OC(═NH)(C₁₋₆alkyl), —OC(═NH)OC₁₋₆ alkyl, —C(═NH)N(C₁₋₆ alkyl)₂, —C(═NH)NH(C₁₋₆alkyl), —C(═NH)NH₂, —OC(═NH)N(C₁₋₆ alkyl)₂, —OC(NH)NH(C₁₋₆ alkyl),—OC(NH)NH₂, —NHC(NH)N(C₁₋₆ alkyl)₂, —NHC(═NH)NH₂, —NHSO₂(C₁₋₆ alkyl),—SO₂N(C₁₋₆ alkyl)₂, —SO₂NH(C₁₋₆ alkyl), —SO₂NH₂, —SO₂C₁₋₆ alkyl,—SO₂OC₁₋₆ alkyl, —OSO₂C₁₋₆ alkyl, —SOC₁₋₆ alkyl, —Si(C₁₋₆ alkyl)₃,—OSi(C₁₋₆ alkyl)₃-C(═S)N(C₁₋₆ alkyl)₂, C(═S)NH(C₁₋₆ alkyl), C(═S)NH₂,—C(═O)S(C₁₋₆ alkyl), —C(═S)SC₁₋₆ alkyl, —SC(═S)SC₁₋₆ alkyl, —P(═O)₂(C₁₋₆alkyl), —P(═O)(C₁₋₆ alkyl)₂, —OP(═O)(C₁₋₆ alkyl)₂, —OP(═O)(OC₁₋₆alkyl)₂, C₁₋₆ alkyl, C₁₋₆ perhaloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₁₀ carbocyclyl, C₆₋₁₀ aryl, 3-10 membered heterocyclyl, 5-10 memberedheteroaryl; or two geminal R^(gg) substituents can be joined to form ═Oor ═S; wherein X is a counterion.

A “counterion” or “anionic counterion” is a negatively charged groupassociated with a cationic quaternary amino group in order to maintainelectronic neutrality. Exemplary counterions include halide ions (e.g.,F⁻, Cl⁻, Br⁻, I⁻), NO₃ ⁻, ClO₄ ⁻, OH⁻, H₂PO₄ ⁻, HSO₄ ⁻, sulfonate ions(e.g., methansulfonate, trifluoromethanesulfonate, p-toluenesulfonate,benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate,naphthalene-1-sulfonic acid-5-sulfonate, ethan-1-sulfonicacid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate,ethanoate, propanoate, benzoate, glycerate, lactate, tartrate,glycolate, and the like).

“Halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro,—Cl), bromine (bromo, —Br), or iodine (iodo, —I).

Nitrogen atoms can be substituted or unsubstituted as valency permits,and include primary, secondary, tertiary, and quaternary nitrogen atoms.Exemplary nitrogen atom substitutents include, but are not limited to,hydrogen, —OH, —OR^(aa), —N(R^(cc))₂, —CN, —C(═O)R^(aa),—C(═O)N(R^(cc))₂, —CO₂R^(aa), —SO₂R^(aa), —C(═NR^(bb))R^(aa),—C(═NR^(cc))OR^(aa), —C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc),—SO₂OR^(cc), —SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc),—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)₂N(R^(cc))₂, —P(═O)(NR^(cc))₂,C₁₋₁₀ alkyl, C₁₋₁₀ perhaloalkyl, C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl, or two R^(cc) groups attached to a nitrogen atom are joinedto form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring,wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl,and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5R^(dd) groups, and wherein R^(aa), R^(bb), R^(cc) and R^(dd) are asdefined above.

In certain embodiments, the substituent present on a nitrogen atom is anitrogen protecting group (also referred to as an amino protectinggroup). Nitrogen protecting groups include, but are not limited to, —OH,—OR^(aa), —N(R^(cc))₂, —C(═O)R^(aa), —C(═O)N(R^(cc))₂, —CO₂R^(aa),—SO₂R^(aa), —C(═NR^(cc))R^(aa), —C(═NR^(cc))OR^(aa),—C(═NR^(cc))N(R^(cc))₂, —SO₂N(R^(cc))₂, —SO₂R^(cc), —SO₂OR^(cc),—SOR^(aa), —C(═S)N(R^(cc))₂, —C(═O)SR^(cc), —C(═S)SR^(cc), C₁₋₁₀ alkyl(e.g., aralkyl, heteroaralkyl), C₂₋₁₀ alkenyl, C₂₋₁₀ alkynyl, C₃₋₁₀carbocyclyl, 3-14 membered heterocyclyl, C₆₋₁₄ aryl, and 5-14 memberedheteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclyl,heterocyclyl, aralkyl, aryl, and heteroaryl is independently substitutedwith 0, 1, 2, 3, 4, or 5 R^(dd) groups, and wherein R^(aa), R^(bb),R^(cc), and R^(dd) are as defined herein. Nitrogen protecting groups arewell known in the art and include those described in detail inProtecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts,3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Amide nitrogen protecting groups (e.g., —C(═O)R^(aa)) include, but arenot limited to, formamide, acetamide, chloroacetamide,trichloroacetamide, trifluoroacetamide, phenylacetamide,3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide,N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide,o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide,(N′-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide,3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide,2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide,3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine,o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

Carbamate nitrogen protecting groups (e.g., —C(═O)OR^(aa)) include, butare not limited to, methyl carbamate, ethyl carbamante,9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate,2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methylcarbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc),2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate(Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate,1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC),1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC),1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc),1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2′- and4′-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamido)ethylcarbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinylcarbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate(Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc),8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz),p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate,2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate,2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methylcarbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc),2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate(Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc),1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate,p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate,2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate,3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzylcarbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentylcarbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate,2,2-dimethoxyacylvinyl carbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethylcarboxamido)propyl carbamate,1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate,2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate,isobutyl carbamate, isonicotinyl carbamate,p-(p′-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate,1-methylcyclohexyl carbamate, 1-methyl-1-cyclopropylmethyl carbamate,1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate,1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate,p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate,4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzylcarbamate.

Sulfonamide nitrogen protecting groups (e.g., —S(═O)₂R^(aa)) include,but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide,2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr),2,4,6-trimethoxybenzenesulfonamide (Mtb),2,6-dimethyl-4-methoxybenzenesulfonamide (Pme),2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte),4-methoxybenzenesulfonamide (Mbs), 2,4,6-trimethylbenzenesulfonamide(Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds),2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide(Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide,4-(4′,8′-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS),benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

Other nitrogen protecting groups include, but are not limited to,phenothiazinyl-(10)-acyl derivative, N′-p-toluenesulfonylaminoacylderivative, N′-phenylaminothioacyl derivative, N-benzoylphenylalanylderivative, N-acetylmethionine derivative,4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts),N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole,N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE),5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted3,5-dinitro-4-pyridone, N-methylamine, N-allylamine,N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine,N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammoniumsalts, N-benzylamine, N-di(4-methoxyphenyl)methylamine,N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr),N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr),N-9-phenylfluorenylamine (PhF),N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm),N-2-picolylamino N′-oxide, N-1,1-dimethylthiomethyleneamine,N-benzylideneamine, N-p-methoxybenzylideneamine,N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine,N—(N′,N′-dimethylaminomethylene)amine, N,N′-isopropylidenediamine,N-p-nitrobenzylideneamine, N-salicylideneamine,N-5-chlorosalicylideneamine,N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine,N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine,N-borane derivative, N-diphenylborinic acid derivative,N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate,N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide,diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt),diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzylphosphoramidate, diphenyl phosphoramidate, benzenesulfenamide,o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide,pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide,triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

In certain embodiments, the substituent present on an oxygen atom is anoxygen protecting group (also referred to as a hydroxyl protectinggroup). Oxygen protecting groups include, but are not limited to,—R^(aa), —N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa),—C(═O)N(R^(bb))₂, —C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa),—C(═NR^(bb))N(R^(bb))₂, —S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃,—P(R^(cc))₂, —P(R^(cc))₃, —P(═O)₂R^(aa), —P(═O)(R^(aa))₂,—P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and —P(═O)(NR^(bb))₂, whereinR^(aa), R^(bb), and R^(cc) are as defined herein. Oxygen protectinggroups are well known in the art and include those described in detailin Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M.Wuts, 3^(rd) edition, John Wiley & Sons, 1999, incorporated herein byreference.

Exemplary oxygen protecting groups include, but are not limited to,methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl,(phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM),p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM),guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM),siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl,bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR),tetrahydropyranyl (THP), 3-bromotetrahydropyranyl,tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl(MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranylS,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl,2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl,1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl,1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl,2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl,t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl,benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl,p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl,p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxido,diphenylmethyl, p,p′-dinitrobenzhydryl, 5-dibenzosuberyl,triphenylmethyl, α-naphthyldiphenylmethyl,p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl,tri(p-methoxyphenyl)methyl, 4-(4′-bromophenacyloxyphenyl)diphenylmethyl,4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl,4,4′,4″-tris(levulinoyloxyphenyl)methyl,4,4′,4″-tris(benzoyloxyphenyl)methyl,3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl,1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl, 9-anthryl,9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl,1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxido, trimethylsilyl(TMS), triethylsilyl (TES), triisopropylsilyl (TIPS),dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS),dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl(TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl,diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate,benzoylformate, acetate, chloroacetate, dichloroacetate,trichloroacetate, trifluoroacetate, methoxyacetate,triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate,3-phenylpropionate, 4-oxopentanoate (levulinate),4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate,adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate,2,4,6-trimethylbenzoate (mesitoate), t-butyl carbonate (BOC), alkylmethyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkyl ethylcarbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc),2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethylcarbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkylisobutyl carbonate, alkyl vinyl carbonate, alkyl allyl carbonate, alkylp-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzylcarbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzylcarbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate,4-ethoxy-1-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate,4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate,2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl,4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate,2,6-dichloro-4-methylphenoxyacetate,2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate,2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate,isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate,o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkylN,N,N′N′-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate,dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate,methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts).

In certain embodiments, the substituent present on a sulfur atom is asulfur protecting group (also referred to as a thiol protecting group).Sulfur protecting groups include, but are not limited to, —R^(aa),—N(R^(bb))₂, —C(═O)SR^(aa), —C(═O)R^(aa), —CO₂R^(aa), —C(═O)N(R^(bb))₂,—C(═NR^(bb))R^(aa), —C(═NR^(bb))OR^(aa), —C(═NR^(bb))N(R^(bb))₂,—S(═O)R^(aa), —SO₂R^(aa), —Si(R^(aa))₃, —P(R^(cc))₂, —P(R^(cc))₃,—P(═O)₂R^(aa), —P(═O)(R^(aa))₂, —P(═O)(OR^(cc))₂, —P(═O)₂N(R^(bb))₂, and—P(═O)(NR^(bb))₂, wherein R^(aa), R^(bb), and R^(cc) are as definedherein. Sulfur protecting groups are well known in the art and includethose described in detail in Protecting Groups in Organic Synthesis, T.W. Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley & Sons, 1999,incorporated herein by reference.

These and other exemplary substituents are described in more detail inthe Detailed Description, Examples, and claims. The present disclosureis not intended to be limited in any manner by the above exemplarylisting of substituents.

“Pharmaceutically acceptable salt” refers to those salts which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and other animals without undue toxicity,irritation, allergic response, and the like, and are commensurate with areasonable benefit/risk ratio. Pharmaceutically acceptable salts arewell known in the art. For example, Berge et al. describepharmaceutically acceptable salts in detail in J. PharmaceuticalSciences (1977) 66:1-19. Pharmaceutically acceptable salts of thecompounds describe herein include those derived from suitable inorganicand organic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid, or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. Representativealkali or alkaline earth metal salts include sodium, lithium, potassium,calcium, magnesium, and the like. Further pharmaceutically acceptablesalts include, when appropriate, quaternary salts.

A “subject” to which administration is contemplated includes, but is notlimited to, humans (e.g., a male or female of any age group, e.g., apediatric subject (e.g, infant, child, adolescent) or adult subject(e.g., young adult, middle-aged adult or senior adult)) and/or othernon-human animals, for example, non-human mammals (e.g., primates (e.g.,cynomolgus monkeys, rhesus monkeys); commercially relevant mammals suchas cattle, pigs, horses, sheep, goats, cats, and/or dogs), birds (e.g.,commercially relevant birds such as chickens, ducks, geese, and/orturkeys), rodents (e.g., rats and/or mice), reptiles, amphibians, andfish. In certain embodiments, the non-human animal is a mammal. Thenon-human animal may be a male or female at any stage of development. Anon-human animal may be a transgenic animal.

“Condition,” “disease,” and “disorder” are used interchangeably herein.

“Treat,” “treating” and “treatment” encompasses an action that occurswhile a subject is suffering from a condition which reduces the severityof the condition or retards or slows the progression of the condition(“therapeutic treatment”). “Treat,” “treating” and “treatment” alsoencompasses an action that occurs before a subject begins to suffer fromthe condition and which inhibits or reduces the severity of thecondition (“prophylactic treatment”).

An “effective amount” of a compound refers to an amount sufficient toelicit the desired biological response, e.g., treat the condition. Aswill be appreciated by those of ordinary skill in this art, theeffective amount of a compound described herein may vary depending onsuch factors as the desired biological endpoint, the pharmacokinetics ofthe compound, the condition being treated, the mode of administration,and the age and health of the subject. An effective amount encompassestherapeutic and prophylactic treatment.

A “therapeutically effective amount” of a compound is an amountsufficient to provide a therapeutic benefit in the treatment of acondition or to delay or minimize one or more symptoms associated withthe condition. A therapeutically effective amount of a compound means anamount of therapeutic agent, alone or in combination with othertherapies, which provides a therapeutic benefit in the treatment of thecondition. The term “therapeutically effective amount” can encompass anamount that improves overall therapy, reduces or avoids symptoms orcauses of the condition, or enhances the therapeutic efficacy of anothertherapeutic agent.

A “prophylactically effective amount” of a compound is an amountsufficient to prevent a condition, or one or more symptoms associatedwith the condition or prevent its recurrence. A prophylacticallyeffective amount of a compound means an amount of a therapeutic agent,alone or in combination with other agents, which provides a prophylacticbenefit in the prevention of the condition. The term “prophylacticallyeffective amount” can encompass an amount that improves overallprophylaxis or enhances the prophylactic efficacy of anotherprophylactic agent.

As used herein, the term “methyltransferase” represents transferaseclass enzymes that are able to transfer a methyl group from a donormolecule to an acceptor molecule, e.g., an amino acid residue of aprotein or a nucleic base of a DNA molecule. Methytransferases typicallyuse a reactive methyl group bound to sulfur in S-adenosyl methionine(SAM) as the methyl donor. In some embodiments, a methyltransferasedescribed herein is a protein methyltransferase. In some embodiments, amethyltransferase described herein is a histone methyltransferase.Histone methyltransferases (HMT) are histone-modifying enzymes,(including histone-lysine N-methyltransferase and histone-arginineN-methyltransferase), that catalyze the transfer of one or more methylgroups to lysine and arginine residues of histone proteins. In certainembodiments, a methyltransferase described herein is a histone-arginineN-methyltransferase.

As generally described above, provided herein are compounds useful asarginine methyltransferase (RMT) inhibitors. In some embodiments, thepresent disclosure provides a compound of Formula (I):

or a pharmaceutically acceptable salt thereof,wherein:

X is N, Z is NR⁴, and Y is CR⁵; or

X is NR⁴, Z is N, and Y is CR⁵; or

X is CR⁵, Z is NR⁴, and Y is N; or

X is CR⁵, Z is N, and Y is NR⁴;

R^(x) is optionally substituted C₁₋₄ alkyl or optionally substitutedC₃₋₄ cycloalkyl;

L₁ is a bond, —O—, —N(R^(B))—, —S—, —C(O)—, —C(O)O—, —C(O)S—,—C(O)N(R^(B))—, —C(O)N(R^(B))N(R^(B))—, —OC(O)—, —OC(O)N(R^(B))—,—NR^(B)C(O)—, —NR^(B)C(O)N(R^(B))—, —NR^(B)C(O)N(R^(B))N(R^(B))—,—NR^(B)C(O)O—, —SC(O)—, —C(═NR^(B))—, —C(═NNR^(B))—, —C(═NOR^(A))—,—C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—, —C(S)—, —C(S)N(R^(B)),—NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—, —N(R^(B))SO₂—,—SO₂N(R^(B))—, or an optionally substituted C₁₋₆ saturated orunsaturated hydrocarbon chain, wherein one or more methylene units ofthe hydrocarbon chain is optionally and independently replaced with —O—,—N(R^(B))—, —S—, —C(O)—, —C(O)O—, —C(O)S—, —C(O)N(R^(B))—,—C(O)N(R^(B))N(R^(B))—, —OC(O)—, —OC(O)N(R^(B))—, —NR^(B)C(O)—,—NR^(B)C(O)N(R^(B))—, —NR^(B)C(O)N(R^(B))N(R^(B))—, —NR^(B)C(O)O—,—SC(O)—, —C(═NR^(B))—, —C(═NNR^(B))—, —C(═NOR^(A))—,—C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—, —C(S)—, —C(S)N(R^(B))—,—NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—, —N(R^(B))SO₂—, or—SO₂N(R^(B))—;

each R^(A) is independently selected from the group consisting ofhydrogen, optionally substituted alkyl, optionally substituted alkenyl,optionally substituted alkynyl, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl,optionally substituted heteroaryl, an oxygen protecting group whenattached to an oxygen atom, and a sulfur protecting group when attachedto a sulfur atom;

each R^(B) is independently selected from the group consisting ofhydrogen, optionally substituted alkyl, optionally substituted alkenyl,optionally substituted alkynyl, optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl,optionally substituted heteroaryl, and a nitrogen protecting group, oran R^(B) and R^(W) on the same nitrogen atom may be taken together withthe intervening nitrogen to form an optionally substituted heterocyclicring;

R^(W) is hydrogen, halogen, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, or optionally substituted heteroaryl, provided thatwhen L₁ is a bond, R^(W) is not optionally substituted aryl oroptionally substituted heteroaryl;

R³ is hydrogen, C₁₋₄ alkyl, or C₃₋₄ cycloalkyl;

R⁴ is hydrogen, optionally substituted C₁₋₆ alkyl, optionallysubstituted C₂₋₆ alkenyl, optionally substituted C₂₋₆ alkynyl,optionally substituted carbocyclyl, optionally substituted heterocyclyl;or optionally substituted C₁₋₄ alkyl-Cy;

Cy is optionally substituted C₃₋₇ cycloalkyl, optionally substituted4-to 7-membered heterocyclyl, optionally substituted aryl, or optionallysubstituted heteroaryl; and

R⁵ is hydrogen, halo, —CN, optionally substituted C₁₋₄ alkyl, oroptionally substituted C₃₋₄ cycloalkyl.

In certain embodiments, R⁴ is hydrogen, optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆ alkenyl, optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₇ cycloalkyl, optionally substituted4-to 7-membered heterocyclyl; or optionally substituted C₁₋₄ alkyl-Cy.

In certain embodiments, when L₁ is a bond, then R^(W) is not hydrogen.In certain embodiments, when L₁ is a bond, R^(W) is not hydrogen,optionally substituted aryl, or optionally substituted heteroaryl.

However, in certain embodiments, L₁ is a bond and R^(W) is hydrogen,halogen, or optionally substituted C₁₋₆alkyl. In certain embodiments, L₁is a bond, R^(W) is hydrogen, halogen, or optionally substitutedC₁₋₆alkyl, X is CR⁵, Z is N, and Y is NR⁴, wherein R⁴ is optionallysubstituted carbocyclyl or optionally substituted heterocyclyl, and insuch instances R⁴ is also referred to as Ring A.

As generally described herein, R^(W) may also be referred to as Ring A,wherein Ring A is optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, or optionallysubstituted heteroaryl, provided that when L₁ is a bond, R^(W) is notoptionally substituted aryl or optionally substituted heteroaryl. R^(W)and Ring A are thus used interchangeably herein when R^(W) is describesa cyclic moiety. Furthermore, as described above, in certainembodiments, Ring A and R⁴ are used interchangeably herein when R⁴encompass an optionally substituted carbocyclyl or optionallysubstituted heterocyclyl group.

In certain embodiments, a provided compound is of Formula (II):

or a pharmaceutically acceptable salt thereof, wherein R^(W), L₁, R³,R⁴, R⁵, and R^(x) are as described herein. In certain embodiments, R^(x)is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R⁴ is hydrogen or optionally substituted C₁₋₆ alkyl (e.g.,methyl). In certain embodiments, R⁵ is hydrogen. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, L₁ is a bond and R^(W) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl). Incertain embodiments, L₁ is an optionally substituted C₂₋₆alkylene,optionally substituted C₂₋₆alkenylene, or optionally substitutedC₂₋₆alkynylene chain, and R^(W) is optionally substituted aryl oroptionally substituted heteroaryl.

In certain embodiments, a provided compound is of Formula (III):

or a pharmaceutically acceptable salt thereof, wherein R^(W), L₁, R³,R⁴, R⁵, and R^(x) are as described herein. In certain embodiments, R^(x)is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R⁴ is hydrogen or optionally substituted C₁₋₆ alkyl (e.g.,methyl). In certain embodiments, R⁵ is hydrogen. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, L₁ is a bond and R^(W) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl). Incertain embodiments, L₁ is an optionally substituted C₂₋₆alkylene,optionally substituted C₂₋₆alkenylene, or optionally substitutedC₂₋₆alkynylene chain, and R^(W) is optionally substituted aryl oroptionally substituted heteroaryl.

In certain embodiments, a provided compound is of Formula (IV):

or a pharmaceutically acceptable salt thereof, wherein R^(W), L₁, R³,R⁴, R⁵, and R^(x) are as described herein. In certain embodiments, R^(x)is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R⁴ is hydrogen or optionally substituted C₁₋₆ alkyl (e.g.,methyl). In certain embodiments, R⁵ is hydrogen. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, L₁ is a bond and R^(W) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl). Incertain embodiments, L₁ is an optionally substituted C₂₋₆alkylene,optionally substituted C₂₋₆alkenylene, or optionally substitutedC₂₋₆alkynylene chain, and R^(W) is optionally substituted aryl oroptionally substituted heteroaryl.

In certain embodiments, a provided compound is of Formula (V):

or a pharmaceutically acceptable salt thereof, wherein R^(W), L₁, R³,R⁴, R⁵, and R^(x) are as described herein. In certain embodiments, R^(x)is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R⁴ is hydrogen or optionally substituted C₁₋₆ alkyl (e.g.,methyl). In certain embodiments, R⁵ is hydrogen. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, L₁ is a bond and R^(W) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl). Incertain embodiments, L₁ is an optionally substituted C₂₋₆alkylene,optionally substituted C₂₋₆alkenylene, or optionally substitutedC₂₋₆alkynylene chain, and R^(W) is optionally substituted aryl oroptionally substituted heteroaryl. In certain embodiments, L₁ is a bond,R^(W) is hydrogen, and R⁴ is an optionally substituted carbocyclyl(e.g., an optionally substituted spiro-fused bicyclic carbocyclyl) oroptionally substituted heterocyclyl (e.g., optionally substitutedspiro-fused bicyclic heterocyclyl).

In certain embodiments, when R⁴ is Ring A, wherein Ring A is anoptionally substituted carbocyclyl or optionally substitutedheterocyclyl, a provided compound is of Formula (XII-a5):

or a pharmaceutically acceptable salt thereof, wherein R^(W), L₁, R³,R⁵, R^(x), and Ring A are as described herein. In certain embodiments,R^(x) is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R⁵ is hydrogen. In certain embodiments, L₁ is a bond, R^(W)is hydrogen, halogen, or optionally substituted C₁₋₄alkyl. In certainembodiments, L₁ is a bond and R^(W) is hydrogen. In certain embodiments,L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. In certainembodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro, chloro,bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W) isoptionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl, e.g.,methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl. Incertain embodiments, Ring A is an optionally substituted bicycliccarbocyclyl (e.g., an optionally substituted spiro-fused bicycliccarbocyclyl) or optionally substituted bicyclic heterocyclyl (e.g.,optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI):

or a pharmaceutically acceptable salt thereof, wherein L₁, Ring A, R³,R⁴, R⁵, and R^(x) are as described herein. In certain embodiments, R^(x)is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R⁴ is hydrogen or optionally substituted C₁₋₆ alkyl (e.g.,methyl). In certain embodiments, R⁵ is hydrogen. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, L₁ is a bond and R^(W) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl). Incertain embodiments, L₁ is an optionally substituted C₂₋₆alkylene,optionally substituted C₂₋₆alkenylene, or optionally substitutedC₂₋₆alkynylene chain, and Ring A (a subset of R^(W)) is optionallysubstituted aryl or optionally substituted heteroaryl.

In certain embodiments, a provided compound is of Formula (VI-a):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R³, R⁴,R⁵, and R^(x) are as described herein. In certain embodiments, R^(x) isoptionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-b):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R³, R⁴,R⁵, and R^(x) are as described herein. In certain embodiments, R^(x) isoptionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-c) or(VI-c′):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R³, R⁴,R⁵, and R^(x) are as described herein. In certain embodiments, R^(x) isoptionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-d):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R^(B),R³, R⁴, R⁵, and R^(x) are as described herein. In certain embodiments,R^(x) is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted C₃₋₆carbocyclyl or optionally substituted 5-6 membered heterocyclyl Incertain embodiments, Ring A (a subset of R^(W)) is an optionallysubstituted bicyclic carbocyclyl (e.g., an optionally substitutedspiro-fused bicyclic carbocyclyl) or optionally substituted bicyclicheterocyclyl (e.g., optionally substituted spiro-fused bicyclicheterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-e):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R^(B),R³, R⁴, R⁵, and R^(x) are as described herein. In certain embodiments,R^(x) is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-f):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R³, R⁴,R⁵, and R^(x) are as described herein. In certain embodiments, R^(x) isoptionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-g):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R^(B),R³, R⁴, R⁵, and R^(x) are as described herein. In certain embodiments,R^(x) is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-h):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R^(B),R³, R⁴, R⁵, and R^(x) are as described herein. In certain embodiments,R^(x) is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-i):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R³, R⁴,R⁵, and R^(x) are as described herein, and p is 1, 2, 3, 4, 5, or 6. Incertain embodiments, R^(x) is optionally substituted C₁₋₄ alkyl (e.g.,methyl). In certain embodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g.,methyl). In certain embodiments, X is N, Z is NR⁴, and Y is CR⁵. Incertain embodiments, X is NR⁴, Z is N, and Y is CR⁵. In certainembodiments, X is CR⁵, Z is NR⁴, and Y is N. In certain embodiments, Xis CR⁵, Z is N, and Y is NR⁴. In certain embodiments, R⁴ is hydrogen oroptionally substituted C₁₋₆ alkyl (e.g., methyl). In certainembodiments, R⁵ is hydrogen. In certain embodiments, Ring A (a subset ofR^(W)) is optionally substituted aryl or optionally substitutedheteroaryl. In certain embodiments, Ring A (a subset of R^(W)) isoptionally substituted C₃₋₆ carbocyclyl or optionally substituted 5-6membered heterocyclyl. In certain embodiments, Ring A (a subset ofR^(W)) is an optionally substituted bicyclic carbocyclyl (e.g., anoptionally substituted spiro-fused bicyclic carbocyclyl) or optionallysubstituted bicyclic heterocyclyl (e.g., optionally substitutedspiro-fused bicyclic heterocyclyl). In certain embodiments, p is 1, 2,or 3.

In certain embodiments, a provided compound is of Formula (VI-j):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R³, R⁴,R⁵, and R^(x) are as described herein. In certain embodiments, R^(x) isoptionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl In certain embodiments,Ring A (a subset of R^(W)) is an optionally substituted bicycliccarbocyclyl (e.g., an optionally substituted spiro-fused bicycliccarbocyclyl) or optionally substituted bicyclic heterocyclyl (e.g.,optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-k):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R^(B),R³, R⁴, R⁵, and R^(x) are as described herein. In certain embodiments,R^(x) is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

In certain embodiments, a provided compound is of Formula (VI-i):

or a pharmaceutically acceptable salt thereof, wherein Ring A, R^(B),R³, R⁴, R⁵, and R^(x) are as described herein. In certain embodiments,R^(x) is optionally substituted C₁₋₄ alkyl (e.g., methyl). In certainembodiments, R³ is hydrogen or C₁₋₄ alkyl (e.g., methyl). In certainembodiments, X is N, Z is NR⁴, and Y is CR⁵. In certain embodiments, Xis NR⁴, Z is N, and Y is CR⁵. In certain embodiments, X is CR⁵, Z isNR⁴, and Y is N. In certain embodiments, X is CR⁵, Z is N, and Y is NR⁴.In certain embodiments, R⁴ is hydrogen or optionally substituted C₁₋₆alkyl (e.g., methyl). In certain embodiments, R⁵ is hydrogen. In certainembodiments, Ring A (a subset of R^(W)) is optionally substituted arylor optionally substituted heteroaryl. In certain embodiments, Ring A (asubset of R^(W)) is optionally substituted C₃₋₆ carbocyclyl oroptionally substituted 5-6 membered heterocyclyl. In certainembodiments, Ring A (a subset of R^(W)) is an optionally substitutedbicyclic carbocyclyl (e.g., an optionally substituted spiro-fusedbicyclic carbocyclyl) or optionally substituted bicyclic heterocyclyl(e.g., optionally substituted spiro-fused bicyclic heterocyclyl).

As defined generally above, L₁ is a bond, —O—, —N(R^(B))—, —S—, —C(O)—,—C(O)O—, —C(O)S—, —C(O)N(R^(B))—, —C(O)N(R^(B))N(R^(B))—, —OC(O)—,—OC(O)N(R^(B))—, —NR^(B)C(O)—, —NR^(B)C(O)N(R^(B))—,—NR^(B)C(O)N(R^(B))N(R^(B))—, —NR^(B)C(O)O—, —SC(O)—, —C(═NR^(B))—,—C(═NNR^(B))—, —C(═NOR^(A))—, —C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—,—C(S)—, —C(S)N(R^(B))—, —NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—,—N(R^(B))SO₂—, —SO₂N(R^(B))—, or an optionally substituted C₁₋₆saturated or unsaturated hydrocarbon chain, wherein one or moremethylene units of the hydrocarbon chain is optionally and independentlyreplaced with —O—, —N(R^(B))—, —S—, —C(O)—, —C(O)O—, —C(O)S—,—C(O)N(R^(B))—, —C(O)N(R^(B))N(R^(B))—, —OC(O)—, —OC(O)N(R^(B))—,—NR^(B)C(O)—, —NR^(B)C(O)N(R^(B))—, —NR^(B)C(O)N(R^(B))N(R^(B))—,—NR^(B)C(O)O—, —SC(O)—, —C(═NR^(B))—, —C(═NNR^(B))—, —C(═NOR^(A))—,—C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—, —C(S)—, —C(S)N(R^(B))—,—NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—, —N(R^(B))SO₂—, or—SO₂N(R^(B))—. In some embodiments, L₁ is a bond. In some embodiments,L₁ is —O—, —N(R^(B))—, —S—. In some embodiments, L₁ is —O—. In someembodiments, L₁ is —N(R^(B))—. In some embodiments, L₁ is —NH_. In someembodiments, L₁ is —C(O)—. In some embodiments, L₁ is —C(O)N(R^(B))— or—NR^(B)C(O)—. In some embodiments, L₁ is —C(O)NH—. In some embodiments,L₁ is —NHC(O)—. In some embodiments, L₁ is —N(R^(B))SO₂— or—SO₂N(R^(B))—. In some embodiments, L₁ is —NHSO₂—. In some embodiments,L₁ is —SO₂NH—.

In some embodiments, L₁ is an optionally substituted C₁₋₆ saturated orunsaturated hydrocarbon chain, wherein one or more methylene units ofthe hydrocarbon chain is optionally and independently replaced with —O—,—N(R^(B))—, —S—, —C(O)—, —C(O)O—, —C(O)S—, —C(O)N(R^(B))—,—C(O)N(R^(B))N(R^(B))—, —OC(O)—, —OC(O)N(R^(B))—, —NR^(B)C(O)—,—NR^(B)C(O)N(R^(B))—, —NR^(B)C(O)N(R^(B))N(R^(B))—, —NR^(B)C(O)O—,—SC(O)—, —C(═NR^(B))—, —C(═NNR^(B))—, —C(═NOR^(A))—,—C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—, —C(S)—, —C(S)N(R^(B))—,—NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—, —N(R^(B))SO₂—, or—SO₂N(R^(B))—. In some embodiments, L₁ is an optionally substituted C₁₋₆saturated or unsaturated hydrocarbon chain. In some embodiments, L₁ isan optionally substituted C₁₋₆ alkylene chain. In some embodiments, L₁is an unsubstituted C₁₋₆ alkylene chain. In some embodiments, L₁ is anoptionally substituted C₂₋₆ alkenylene chain. In some embodiments, L₁ isan unsubstituted C₂₋₆ alkenylene chain. In some embodiments, L₁ is—CH═CH—. In some embodiments, L₁ is an optionally substituted C₂₋₆alkynylene chain. In some embodiments, L₁ is an unsubstituted C₂₋₆alkynylene chain. In some embodiments, L₁ is —C≡C—. In some embodiments,L₁ is an optionally substituted C₁₋₆ saturated or unsaturatedhydrocarbon chain, wherein one methylene unit of the hydrocarbon chainis optionally and independently replaced with —O—, —N(R^(B))—, —S—,—C(O)—, —C(O)O—, —C(O)S—, —C(O)N(R^(B))—, —C(O)N(R^(B))N(R^(B))—,—OC(O)—, —OC(O)N(R^(B))—, —NR^(B)C(O)—, —NR^(B)C(O)N(R^(B))—,—NR^(B)C(O)N(R^(B))N(R^(B))—, —NR^(B)C(O)O—, —SC(O)—, —C(═NR^(B))—,—C(═NNR^(B))—, —C(═NOR^(A))—, —C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—,—C(S)—, —C(S)N(R^(B))—, —NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—,—N(R^(B))SO₂—, or —SO₂N(R^(B))—. In some embodiments, L₁ is anoptionally substituted C₁₋₆ saturated or unsaturated hydrocarbon chain,wherein two methylene units of the hydrocarbon chain is optionally andindependently replaced with —O—, —N(R^(B))—, —S—, —C(O)—, —C(O)O—,—C(O)S—, —C(O)N(R^(B))—, —C(O)N(R^(B))N(R^(B))—, —OC(O)—,—OC(O)N(R^(B))—, —NR^(B)C(O)—, —NR^(B)C(O)N(R^(B))—,—NR^(B)C(O)N(R^(B))N(R^(B))—, —NR^(B)C(O)O—, —SC(O)—, —C(═NR^(B))—,—C(═NNR^(B))—, —C(═NOR^(A))—, —C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—,—C(S)—, —C(S)N(R^(B))—, —NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—,—N(R^(B))SO₂—, or —SO₂N(R^(B))—. In some embodiments, L₁ is anoptionally substituted C₁₋₆ saturated or unsaturated hydrocarbon chain,wherein three methylene units of the hydrocarbon chain is optionally andindependently replaced with —O—, —N(R^(B))—, —S—, —C(O)—, —C(O)O—,—C(O)S—, —C(O)N(R^(B))—, —C(O)N(R^(B))N(R^(B))—, —OC(O)—,—OC(O)N(R^(B))—, —NR^(B)C(O)—, —NR^(B)C(O)N(R^(B))—,—NR^(B)C(O)N(R^(B))N(R^(B))—, —NR^(B)C(O)O—, —SC(O)—, —C(═NR^(B))—,—C(═NNR^(B))—, —C(═NOR^(A))—, —C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—,—C(S)—, —C(S)N(R^(B))—, —NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—,—N(R^(B))SO₂—, or —SO₂N(R^(B))—. In some embodiments, L₁ is anoptionally substituted C₁₋₆ saturated or unsaturated hydrocarbon chain,wherein one or more methylene units of the hydrocarbon chain isoptionally and independently replaced with —O—, —N(R^(B))—, —S—, —C(O)—,—C(O)N(R^(B))—, —NR^(B)C(O)—, —SO₂—, —N(R^(B))SO₂—, or —SO₂N(R^(B))—. Insome embodiments, L₁ is —CH₂O— or —OCH₂—.

As defined generally above, R^(W) is hydrogen, optionally substitutedalkyl, optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, or optionally substituted heteroaryl;provided that when L₁ is a bond, R^(W) is not optionally substitutedaryl or optionally substituted heteroaryl. In certain embodiments, whenL₁ is a bond, then R^(W) is not hydrogen. In some embodiments, when L₁is a bond, R^(W) is not hydrogen, optionally substituted aryl, oroptionally substituted heteroaryl.

In certain embodiments, L₁ is a bond, and R^(W) is hydrogen, halogen, oroptionally substituted C₁₋₆alkyl. In certain embodiments, L₁ is a bondand R^(W) is hydrogen. In certain embodiments, L₁ is a bond and R^(W) ishalogen, e.g., fluoro, chloro, bromo, or iodo. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl, e.g.,unsubstituted C₁₋₆alkyl, e.g., methyl, ethyl, propyl, butyl, isopropyl,isobutyl, or isoamyl. In certain embodiments, L₁ is a bond, R^(W) ishydrogen, halogen, or optionally substituted C₁₋₆alkyl, X is CR⁵, Z isN, and Y is NR⁴, and R⁴ is optionally substituted carbocyclyl oroptionally substituted heterocyclyl (in such instances, R⁴ may also bereferred to as Ring A).

In some embodiments, R^(W) is hydrogen.

In some embodiments, R^(W) is halogen, e.g., fluoro, chloro, bromo, oriodo.

In some embodiments, R^(W) is optionally substituted C₁₋₆ alkyl. In someembodiments, R^(W) is unsubstituted C₁₋₆ alkyl. In some embodiments,R^(W) is methyl, ethyl, propyl, or butyl. In some embodiments, R^(W) isisopropyl, isobutyl, or isoamyl. In some embodiments, R^(W) isoptionally substituted C₂₋₆ alkenyl. In some embodiments, R^(W) is C₂₋₆alkynyl.

In some embodiments, R^(W) is optionally substituted carbocyclyl. Insome embodiments, R^(W) is optionally substituted C₃₋₆ carbocyclyl. Insome embodiments, R^(W) is unsubstituted cyclopropyl. In someembodiments, R^(W) is substituted cyclopropyl. In some embodiments,R^(W) is unsubstituted cyclobutyl. In some embodiments, R^(W) issubstituted cyclobutyl. In some embodiments, R^(W) is unsubstitutedcyclopentyl. In some embodiments, R^(W) is substituted cyclopentyl. Insome embodiments, R^(W) is unsubstituted cyclohexyl. In someembodiments, R^(W) is substituted cyclohexyl. In some embodiments, R^(W)is optionally substituted cyclopentenyl or optionally substitutedcyclohexenyl.

In some embodiments, R^(W) is optionally substituted heterocyclyl. Insome embodiments, R^(W) is an optionally substituted 4-to 7-memberedheterocyclyl ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, R^(W) is azetidinylor oxetanyl. In some embodiments, R^(W) is optionally substitutedtetrahydrofuranyl, optionally substituted pyrrolidinyl, optionallysubstituted dihydropyrrolyl, or optionally substitutedpyrrolyl-2,5-dione. In some embodiments, R^(W) is optionally substitutedpiperidinyl, optionally substituted tetrahydropyranyl, optionallysubstituted dihydropyranyl, optionally substituted dihydropyridinyl, andoptionally substituted thianyl. In certain embodiments, R^(W) isoptionally substituted piperidinyl. In some embodiments, R^(W) isoptionally substituted piperazinyl, optionally substituted morpholinyl,optionally substituted dithianyl, and optionally substituted dioxanyl.In some embodiments, R^(W) is a 5- or 6-membered heterocyclyl groupfused to a C₆ aryl ring. In some embodiments, R^(W) is optionallysubstituted indolinyl, optionally substituted isoindolinyl, optionallysubstituted dihydrobenzofuranyl, optionally substituteddihydrobenzothienyl, or optionally substituted benzoxazolinonyl. In someembodiments, R^(W) is optionally substituted tetrahydroquinolinyl oroptionally substituted tetrahydroisoquinolinyl.

In some embodiments, R^(W) is optionally substituted aryl. In someembodiments, R^(W) is optionally substituted phenyl. In someembodiments, R^(W) is unsubstituted phenyl. In some embodiments, R^(W)is substituted phenyl. In some embodiments, R^(W) is monosubstitutedphenyl. In some embodiments, R^(W) is disubstituted phenyl. In someembodiments, R^(W) is trisubstituted phenyl. In some embodiments, R^(W)is optionally substituted naphthyl. In some embodiments, R^(W) isunsubstituted naphthyl.

In some embodiments, R^(W) is optionally substituted heteroaryl. In someembodiments, R^(W) is an optionally substituted 5-to 10-memberedheteroaryl having 1-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, R^(W) is an optionallysubstituted 5-to 8-membered heteroaryl having 1-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, R^(W) is an optionally substituted 5-to 6-memberedheteroaryl having 1-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, R^(W) is an optionallysubstituted 5-to 6-membered heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, R^(W) is an optionally substituted 5-to 6-memberedheteroaryl having 1-2 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, R^(W) is an optionallysubstituted 5-to 6-membered heteroaryl having 1 heteroatom selected fromnitrogen, oxygen, and sulfur. In some embodiments, R^(W) is anoptionally substituted 8-to 10-membered bicyclic heteroaryl having 1-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Insome embodiments, R^(W) is an optionally substituted 9-to 10-memberedbicyclic heteroaryl having 1-4 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, R^(W) is anoptionally substituted 9-membered bicyclic heteroaryl having 1-4heteroatoms independently selected from nitrogen, oxygen, and sulfur. Insome embodiments, R^(W) is an optionally substituted 9-membered bicyclicheteroaryl having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, R^(W) is an optionallysubstituted 9-membered bicyclic heteroaryl having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, R^(W) is an optionally substituted 9-membered bicyclicheteroaryl having 1 heteroatom selected from nitrogen, oxygen, andsulfur. In some embodiments, R^(W) is an optionally substituted10-membered bicyclic heteroaryl having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In some embodiments, R^(W)is an optionally substituted 10-membered bicyclic heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur. Insome embodiments, R^(W) is an optionally substituted 10-memberedbicyclic heteroaryl having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, R^(W) is anoptionally substituted 10-membered bicyclic heteroaryl having 1heteroatom selected from nitrogen, oxygen, and sulfur.

In some embodiments, R^(W) is substituted or unsubstituted pyrrolyl,substituted or unsubstituted furanyl, substituted or unsubstitutedthienyl, substituted or unsubstituted imidazolyl, substituted orunsubstituted pyrazolyl, substituted or unsubstituted oxazolyl,substituted or unsubstituted thiazolyl, substituted or unsubstitutedisothiazolyl, substituted or unsubstituted triazolyl, substituted orunsubstituted thiadiazolyl, substituted or unsubstituted thiadiazolyl,substituted or unsubstituted tetrazolyl, substituted or unsubstitutedpyridyl, substituted or unsubstituted pyrimidyl, substituted orunsubstituted pyrazinyl, substituted or unsubstituted pyridazinyl,substituted or unsubstituted triazinyl, substituted or unsubstitutedindolyl, substituted or unsubstituted isoindolyl, substituted orunsubstituted indazolyl, substituted or unsubstituted benzotriazolyl,substituted or unsubstituted benzothiophenyl, substituted orunsubstituted isobenzothiophenyl, substituted or unsubstitutedbenzofuranyl, substituted or unsubstituted benzoisofuranyl, substitutedor unsubstituted benzimidazolyl, substituted or unsubstitutedbenzoxazolyl, substituted or unsubstituted benzoxadiazolyl, substitutedor unsubstituted benzisoxazolyl, substituted or unsubstitutedbenzthiazolyl, substituted or unsubstituted benzisothiazolyl,substituted or unsubstituted benzthiadiazolyl, substituted orunsubstituted indolizinyl, substituted or unsubstituted purinyl,substituted or unsubstituted pyrrolopyridinyl, substituted orunsubstituted triazolopyridinyl, substituted or unsubstitutednaphthyridinyl, substituted or unsubstituted pteridinyl, substituted orunsubstituted quinolinyl, substituted or unsubstituted isoquinolinyl,substituted or unsubstituted cinnolinyl, substituted or unsubstitutedquinoxalinyl, substituted or unsubstituted quinazolinyl, or substitutedor unsubstituted phthalazinyl.

As generally described herein, R^(W) may also be referred to as Ring A,wherein Ring A is optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, or optionallysubstituted heteroaryl, provided that when L₁ is a bond, R^(W) is notoptionally substituted aryl or optionally substituted heteroaryl. R^(W)and Ring A are thus used interchangeably herein when R^(W) is describesa cyclic moiety. Furthermore, as described above, in certainembodiments, Ring A and R⁴ are used interchangeably herein when R⁴encompass an optionally substituted carbocyclyl or optionallysubstituted heterocyclyl group.

In some embodiments, Ring A (corresponding to R^(W) or R⁴) is optionallysubstituted carbocyclyl. In some embodiments, Ring A is optionallysubstituted C₃₋₆ carbocyclyl. In some embodiments, Ring A is optionallysubstituted C₃₋₈ carbocyclyl. In some embodiments, Ring A is optionallysubstituted C₃ carbocyclyl, C₄ carbocyclyl, C₅ carbocyclyl, C₆carbocyclyl, C₇ carbocyclyl, or C₈ carbocyclyl. In some embodiments,Ring A is unsubstituted cyclopropyl. In some embodiments, Ring A issubstituted cyclopropyl. In some embodiments, Ring A is unsubstitutedcyclobutyl. In some embodiments, Ring A is substituted cyclobutyl. Insome embodiments, Ring A is unsubstituted cyclopentyl. In someembodiments, Ring A is substituted cyclopentyl. In some embodiments,Ring A is unsubstituted cyclohexyl. In some embodiments, Ring A issubstituted cyclohexyl. In some embodiments, Ring A is optionallysubstituted cyclopentenyl or optionally substituted cyclohexenyl.

In some embodiments, Ring A (corresponding to R^(W) or R⁴) is optionallysubstituted bicyclic carbocyclyl. In certain embodiments, Ring A is afused bicyclic carbocyclyl, e.g., Ring A is an optionally substitutedC₃₋₁₀ carbocyclyl radical comprising an optionally substituted C₃₋₁₀carbocyclyl, optionally substituted aryl, or optionally substitutedheteroaryl ring fused thereto. In certain embodiments, Ring A is abridged bicyclic carbocyclyl, e.g., an optionally substituted C₃₋₁₀carbocyclyl radical bridged by a group comprising 1, 2, 3, 4, or 5linear atoms. In certain embodiments, Ring A is a spiro-fused bicycliccarbocyclyl, e.g., an optionally substituted C₃₋₁₀ carbocyclyl radicalcomprising an optionally substituted C₃₋₁₀ carbocyclyl ring spiro-fusedthereto.

In some embodiments, Ring A (corresponding to R^(W) or R⁴) is optionallysubstituted heterocyclyl. In some embodiments, Ring A is an optionallysubstituted 4-to 7-membered heterocyclyl ring having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, Ring A is azetidinyl or oxetanyl. In some embodiments, RingA is optionally substituted tetrahydrofuranyl, optionally substitutedpyrrolidinyl, optionally substituted dihydropyrrolyl, or optionallysubstituted pyrrolyl-2,5-dione. In some embodiments, Ring A isoptionally substituted piperidinyl, optionally substitutedtetrahydropyranyl, optionally substituted dihydropyranyl, optionallysubstituted dihydropyridinyl, and optionally substituted thianyl. Incertain embodiments, Ring A is optionally substituted piperidinyl. Insome embodiments, Ring A is optionally substituted piperazinyl,optionally substituted morpholinyl, optionally substituted dithianyl,and optionally substituted dioxanyl. In some embodiments, Ring A is a 5-or 6-membered heterocyclyl group fused to a C₆ aryl ring. In someembodiments, Ring A is optionally substituted indolinyl, optionallysubstituted isoindolinyl, optionally substituted dihydrobenzofuranyl,optionally substituted dihydrobenzothienyl, or optionally substitutedbenzoxazolinonyl. In some embodiments, Ring A is optionally substitutedtetrahydroquinolinyl or optionally substituted tetrahydroisoquinolinyl.

In some embodiments, Ring A (corresponding to R^(W) or R⁴) is optionallysubstituted bicyclic heterocyclyl. In certain embodiments, Ring A is afused bicyclic heterocyclyl, e.g., Ring A is an optionally substituted3-to 10-membered heterocyclyl radical comprising an optionallysubstituted C₃₋₁₀ carbocyclyl, optionally substituted 3-to 10-memberedheterocyclyl, optionally substituted aryl, or optionally substitutedheteroaryl ring fused thereto. In certain embodiments, Ring A is abridged bicyclic heterocyclyl, e.g., an optionally substituted 3-to10-membered heterocyclyl radical bridged by a group comprising 1, 2, 3,4, or 5 linear atoms. In certain embodiments, Ring A is a spiro-fusedbicyclic heterocyclyl, e.g., an optionally substituted 3-to 10-memberedheterocyclyl radical comprising an optionally substituted C₃₋₁₀carbocyclyl ring or optionally substituted 3-to 10-membered heterocyclylring spiro-fused thereto.

In some embodiments, Ring A (corresponding to R^(W)) is optionallysubstituted aryl. In some embodiments, Ring A is optionally substitutedphenyl. In some embodiments, Ring A is unsubstituted phenyl. In someembodiments, Ring A is substituted phenyl. In some embodiments, Ring Ais monosubstituted phenyl. In some embodiments, Ring A is disubstitutedphenyl. In some embodiments, Ring A is trisubstituted phenyl. In someembodiments, Ring A is optionally substituted naphthyl. In someembodiments, Ring A is unsubstituted naphthyl.

In some embodiments, Ring A (corresponding to R^(W)) is optionallysubstituted heteroaryl. In some embodiments, Ring A is an optionallysubstituted 5-to 10-membered heteroaryl having 1-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, Ring A is an optionally substituted 5-to 8-memberedheteroaryl having 1-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, Ring A is an optionallysubstituted 5-to 6-membered heteroaryl having 1-4 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, Ring A is an optionally substituted 5-to 6-memberedheteroaryl having 1-3 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, Ring A is an optionallysubstituted 5-to 6-membered heteroaryl having 1-2 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, Ring A is an optionally substituted 5-to 6-memberedheteroaryl having 1 heteroatom selected from nitrogen, oxygen, andsulfur. In some embodiments, Ring A is an optionally substituted 8-to10-membered bicyclic heteroaryl having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In some embodiments, Ring Ais an optionally substituted 9-to 10-membered bicyclic heteroaryl having1-4 heteroatoms independently selected from nitrogen, oxygen, andsulfur. In some embodiments, Ring A is an optionally substituted9-membered bicyclic heteroaryl having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In some embodiments, Ring Ais an optionally substituted 9-membered bicyclic heteroaryl having 1-3heteroatoms independently selected from nitrogen, oxygen, and sulfur. Insome embodiments, Ring A is an optionally substituted 9-memberedbicyclic heteroaryl having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In some embodiments, Ring A is anoptionally substituted 9-membered bicyclic heteroaryl having 1heteroatom selected from nitrogen, oxygen, and sulfur. In someembodiments, Ring A is an optionally substituted 10-membered bicyclicheteroaryl having 1-4 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, Ring A is an optionallysubstituted 10-membered bicyclic heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, Ring A is an optionally substituted 10-membered bicyclicheteroaryl having 1-2 heteroatoms independently selected from nitrogen,oxygen, and sulfur. In some embodiments, Ring A is an optionallysubstituted 10-membered bicyclic heteroaryl having 1 heteroatom selectedfrom nitrogen, oxygen, and sulfur.

In some embodiments, Ring A (corresponding to R^(W)) is substituted orunsubstituted pyrrolyl, substituted or unsubstituted furanyl,substituted or unsubstituted thienyl, substituted or unsubstitutedimidazolyl, substituted or unsubstituted pyrazolyl, substituted orunsubstituted oxazolyl, substituted or unsubstituted thiazolyl,substituted or unsubstituted isothiazolyl, substituted or unsubstitutedtriazolyl, substituted or unsubstituted thiadiazolyl, substituted orunsubstituted thiadiazolyl, substituted or unsubstituted tetrazolyl,substituted or unsubstituted pyridyl, substituted or unsubstitutedpyrimidyl, substituted or unsubstituted pyrazinyl, substituted orunsubstituted pyridazinyl, substituted or unsubstituted triazinyl,substituted or unsubstituted indolyl, substituted or unsubstitutedisoindolyl, substituted or unsubstituted indazolyl, substituted orunsubstituted benzotriazolyl, substituted or unsubstitutedbenzothiophenyl, substituted or unsubstituted isobenzothiophenyl,substituted or unsubstituted benzofuranyl, substituted or unsubstitutedbenzoisofuranyl, substituted or unsubstituted benzimidazolyl,substituted or unsubstituted benzoxazolyl, substituted or unsubstitutedbenzoxadiazolyl, substituted or unsubstituted benzisoxazolyl,substituted or unsubstituted benzthiazolyl, substituted or unsubstitutedbenzisothiazolyl, substituted or unsubstituted benzthiadiazolyl,substituted or unsubstituted indolizinyl, substituted or unsubstitutedpurinyl, substituted or unsubstituted pyrrolopyridinyl, substituted orunsubstituted triazolopyridinyl, substituted or unsubstitutednaphthyridinyl, substituted or unsubstituted pteridinyl, substituted orunsubstituted quinolinyl, substituted or unsubstituted isoquinolinyl,substituted or unsubstituted cinnolinyl, substituted or unsubstitutedquinoxalinyl, substituted or unsubstituted quinazolinyl, or substitutedor unsubstituted phthalazinyl.

In certain embodiments, R^(W) (or Ring A) is of Formula (q-1):

In certain embodiments, R^(W) (or Ring A) is of Formula (q-2):

In certain embodiments, R^(W) (or Ring A) is of Formula (q-3):

In certain embodiments, R^(W) (or Ring A) is of Formula (q-4):

As used herein, each instance of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹may independently be O, S, N, NR^(N), C, or CR^(C), as valency permits,wherein at least one of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ is O, S,N, NR^(N), and wherein:

each instance of R^(C) is independently selected from the groupconsisting of hydrogen, halo, —CN, —NO₂, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted aryl,optionally substituted heterocyclyl, optionally substituted heteroaryl,—OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; and

each instance of R^(N) is independently selected from the groupconsisting of hydrogen, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, optionally substituted heteroaryl, —C(═O)R^(A),—C(═O)OR^(A), —C(═O)SR^(A), —C(═O)N(R^(B))₂, —C(═NR^(B))R^(A),—C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —C(═S)R^(A),—C(═S)N(R^(B))₂, —S(═O)R^(A), —SO₂R^(A), —SO₂N(R^(B))₂, and a nitrogenprotecting group.

In certain embodiments, V¹ is O, S, N or NR^(N). In certain embodiments,V¹ is N or NR^(N). In certain embodiments, V¹ is O. In certainembodiments, V¹ is S. In certain embodiments, only one of V¹, V², V³,V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ is selected from the group consisting of O,S, N, and NR^(N). In certain embodiments, only one of V¹, V², V³, V⁴,V⁵, V⁶, V⁷, V⁸, and V⁹ is selected from the group consisting of N andNR^(N). In certain embodiments, only one of V¹, V², V³, V⁴, V⁵, V⁶, V⁷,V⁸, and V⁹ is O. In certain embodiments, only one of V¹, V², V³, V⁴, V⁵,V⁶, V⁷, V⁸, and V⁹ is S. In certain embodiments, only two of V¹, V², V³,V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ are each independently selected from thegroup consisting of O, S, N, and NR^(N). In certain embodiments, onlytwo of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ are each independentlyselected from the group consisting of N and NR^(N). In certainembodiments, only two of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ are eachindependently selected from the group consisting of O, N and NR^(N). Incertain embodiments, only two of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹are each independently selected from the group consisting of S, N andNR^(N). In certain embodiments, only three of V¹, V², V³, V⁴, V⁵, V⁶,V⁷, V⁸, and V⁹ are each independently selected from the group consistingof O, S, N, and NR^(N). In certain embodiments, only three of V¹, V²,V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ are each independently selected from thegroup consisting of N and NR^(N). In certain embodiments, only three ofV¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ are each independently selectedfrom the group consisting of O, N and NR^(N). In certain embodiments,only three of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ are eachindependently selected from the group consisting of S, N and NR^(N). Incertain embodiments, only four of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹are each independently selected from the group consisting of O, S, N,and NR^(N). In certain embodiments, only four of V¹, V², V³, V⁴, V⁵, V⁶,V⁷, V⁸, and V⁹ are each independently selected from the group consistingof N and NR^(N). In certain embodiments, only four of V¹, V², V³, V⁴,V⁵, V⁶, V⁷, V⁸, and V⁹ are each independently selected from the groupconsisting of O, N and NR^(N). In certain embodiments, only four of V¹,V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ are each independently selected fromthe group consisting of S, N and NR^(N). In certain embodiments, onlyfive of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ are each independentlyselected from the group consisting of O, S, N, and NR^(N). In certainembodiments, only five of V¹, V², V³, V⁴, V⁵, V⁶, V⁷, V⁸, and V⁹ areeach independently selected from the group consisting of N and NR^(N).

In certain embodiments, R^(W) (or Ring A) is an optionally substituted5-membered heteroaryl ring. In certain embodiments, R^(W) (or Ring A) isof Formula (q-5):

As used herein, each instance of V¹⁰, V¹¹, V¹², V¹³, and V¹⁴ mayindependently be O, S, N, NR^(N), C, or CR^(C), as valency permits,wherein R^(N) and R^(C) are as defined herein, and wherein at least oneof V¹⁰, V¹¹, V¹², V¹³, and V¹⁴ is O, S, N, or NR^(N). In certainembodiments, only one of V¹⁰, V¹¹, V², V¹³, and V¹⁴ is selected from thegroup consisting of O, S, N, and NR^(N). In certain embodiments, onlytwo of V, V¹¹, V¹², V¹³, and V¹⁴ are selected from the group consistingof O, S, N, and NR^(N). In certain embodiments, only three of V¹⁰, V¹¹,V¹², V¹³, and V¹⁴ are selected from the group consisting of O, S, N, andNR^(N). In certain embodiments, only four of V¹⁰, V¹¹, V¹², V³, and V¹⁴are selected from the group consisting of O, S, N, and NR^(N).

In certain embodiments, R^(W) (or Ring A) is an optionally substituted6-membered heteroaryl ring. In certain embodiments, R^(W) (or Ring A) isof Formula (q-6):

In compounds of Formula (q-6), V¹⁵, V¹⁶, V¹⁷, V¹⁸, V¹⁹, and V²⁰ are eachindependently selected from the group consisting of N or CR^(C), whereinat least one of V¹⁵, V¹⁶, V¹⁷, V¹⁸, V¹⁹, and V²⁰ is N. In certainembodiments, only one of V¹⁵, V¹⁶, V¹⁷, V¹⁸, V¹⁹, and V²⁰ is N. Incertain embodiments, only two of V¹⁵, V¹⁶, V¹⁷, V¹⁸, V¹⁹, and V²⁰ are N.In certain embodiments, only three of V¹⁵, V¹⁶, V¹⁷, V¹⁸, V¹⁹, and V²⁰are N.

In certain embodiments, Ring A (corresponding to R^(W) or R⁴) is anoptionally substituted carbocyclyl or optionally substitutedheterocyclyl, e.g., an optionally substituted C₆ carbocyclyl, optionallysubstituted 6-membered heterocyclyl, optionally substituted C₆carbocyclyl radical comprising an optionally substituted C₃₋₁₀carbocyclyl ring or optionally substituted 3-to 10-membered heterocyclylring spiro-fused thereto, or an optionally substituted 6-memberedheterocyclyl radical comprising an optionally substituted C₃₋₁₀carbocyclyl ring or optionally substituted 3-to 10-membered heterocyclylring spiro-fused thereto. Furthermore, in certain embodiments, L₁ is abond, R^(W) is hydrogen, halogen, or optionally substituted C₁₋₆alkyl, Xis CR⁵, Z is N, and Y is NR⁴, and R⁴ is optionally substitutedcarbocyclyl or optionally substituted heterocyclyl, e.g., an optionallysubstituted C₆ carbocyclyl, optionally substituted 6-memberedheterocyclyl, optionally substituted C₆ carbocyclyl radical comprisingan optionally substituted C₃₋₁₀ carbocyclyl ring or optionallysubstituted 3-to 10-membered heterocyclyl ring spiro-fused thereto, oran optionally substituted 6-membered heterocyclyl radical comprising anoptionally substituted C₃₋₁₀ carbocyclyl ring or optionally substituted3-to 10-membered heterocyclyl ring spiro-fused thereto.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula (q-7)-(q-17):

wherein:

V²¹, V²², V²³, and V²⁴ are each independently O, S, NR^(N), C═O, orC(R^(C))₂ as valency permits, provided no more than two of V²¹, V²²,V²³, and V²⁴ is a heteroatom O, S, and NR^(N), alternatively wherein oneof V²¹, V²², V²³, and V²⁴ and another of V²¹, V²², V²³, and V²⁴ adjacentto each other are joined to form an N═C(R^(C)) or C(R^(C))═C(R^(C))group provided the ring thus formed is not an aromatic ring;

each instance of R^(C) is independently hydrogen, halo, —CN, —NO₂,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted carbocyclyl, optionallysubstituted aryl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂;

each instance of R^(N) is independently hydrogen, optionally substitutedalkyl, optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl,—C(═O)R^(A), —C(═O)OR^(A), —C(═O)SR^(A), —C(═O)N(R^(B))₂,—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —C(═S)R^(A), —C(═S)N(R^(B))₂, —S(═O)R^(A),—SO₂R^(A), —SO₂N(R^(B))₂, or a nitrogen protecting group, or two R^(N)groups are joined to form an optionally substituted heterocyclic ring,or one R^(N) group and one R^(D) group are joined to form an optionallysubstituted heterocyclic ring;

each instance of R^(F) is independently hydrogen or halo; and

each instance of R^(D) is independently hydrogen, halo, —CN, —NO₂,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted carbocyclyl, optionallysubstituted aryl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or two R^(D) groups are joined toform an optionally substituted carbocyclic or optionally substitutedheterocyclic ring.

In certain embodiments, each instance of V²¹, V²², V²³, and V²⁴ isC(R^(C))₂. In certain embodiments, one instance of V²¹, V²², V²³, andV²⁴ is O. In certain embodiments, one instance of V²¹, V²², V²³, and V²⁴is S. In certain embodiments, one instance of V²¹, V²², V²³, and V²⁴ isNR^(N). In certain embodiments, only one of V²¹, V²², V²³, and V²⁴ is aheteroatom selected from O, S, and NR^(N). In certain embodiments, twoof V²¹, V²², V²³, and V²⁴ is a heteroatom selected from O, S, andNR^(N).

In certain embodiments, V²¹ is a heteroatom selected from O, S, andNR^(N). In certain embodiments, V²¹ is O. In certain embodiments, V²¹ isNR^(N). For example, in certain embodiments, wherein V²¹ is O or NR^(N),provided is an Ring A (R^(W) or R⁴) group of Formula:

In certain embodiments, V²² is a heteroatom selected from O, S, andNR^(N). In certain embodiments, V²² is O. In certain embodiments, V²² isNR^(N). For example, in certain embodiments, wherein V²² is O or NR^(N),provided is an Ring A (R^(W) or R⁴) group of Formula:

In certain embodiments, V²³ is a heteroatom selected from O, S, andNR^(N). In certain embodiments, V²³ is O. In certain embodiments, V²³ isNR^(N). For example, in certain embodiments, wherein V²³ is O or NR^(N),provided is an Ring A (R^(W) or R⁴) group of Formula:

In certain embodiments, V²⁴ is a heteroatom selected from O, S, andNR^(N). In certain embodiments, V²⁴ is O. In certain embodiments, V²⁴ isNR^(N). For example, in certain embodiments, wherein V²⁴ is O or NR^(N),provided is Ring A (R^(W) or R⁴) group of Formula:

In certain embodiments, at least one instance of R^(D) is hydrogen,e.g., each instance for (q-10); 1 or 2 instances for (q-7), (q-9),(q-12), (q-13), (q-14), (q-15), (q-16), and (q-17); and 1, 2, 3, or 4instances for (q-8). However, in certain embodiments, at least oneinstance of R^(D) is a non-hydrogen group, e.g., each instance for(q-10); 1 or 2 instances for (q-7), (q-9), (q-12), (q-13), (q-14),(q-15), (q-16), and (q-17); and 1, 2, 3, or 4 instances for (q-8). Forexample, in certain embodiments, each instance of R^(D) is anon-hydrogen group.

In certain embodiments, two R^(D) groups are joined to form anoptionally substituted carbocyclic ring, e.g., an optionally substitutedC₃₋₆ carbocyclic ring. In certain embodiments, two R^(D) groups arejoined to form an optionally substituted C₃ carbocyclic ring, optionallysubstituted C₄ carbocyclic ring, optionally substituted C₅ carbocyclicring, or optionally substituted C₆ carbocyclic ring. In certainembodiments, the R^(D) groups are joined to form an unsubstitutedcarbocyclic ring. However, in certain embodiments, the R^(D) groups arejoined to form a substituted carbocyclic ring, e.g., substituted withone or more alkyl groups.

In certain embodiments, two R^(D) groups are joined to form anoptionally substituted heterocyclic ring, e.g., an optionallysubstituted 3-to 6-membered heterocyclic ring comprising 1 or 2heteroatoms selected from oxygen, nitrogen, or sulfur. In certainembodiments, two R^(D) groups are joined to form an optionallysubstituted 3-membered heterocyclic ring, optionally substituted4-membered heterocyclic ring, optionally substituted 5-memberedheterocyclic ring, or optionally substituted 6-membered heterocyclicring. In certain embodiments, the R^(D) groups are joined to form anunsubstituted heterocyclic ring. However, in certain embodiments, theR^(D) groups are joined to form a substituted heterocyclic ring, e.g.,substituted with one or more alkyl groups.

In certain embodiments, two R^(N) groups are joined to form anoptionally substituted heterocyclic ring, e.g., an optionallysubstituted 3-to 6-membered heterocyclic ring. In certain embodiments,two R^(N) groups are joined to form an optionally substituted 5-memberedheterocyclic ring, or optionally substituted 6-membered heterocyclicring. In certain embodiments, the R^(N) groups are joined to form anunsubstituted heterocyclic ring. However, in certain embodiments, theR^(N) groups are joined to form a substituted heterocyclic ring, e.g.,substituted with one or more alkyl groups.

In certain embodiments, one R^(N) group and one R^(D) group (e.g.,wherein the N atom to which the R^(N) group is attached and the R^(D)group are joined to the same carbon atom) are joined to form anoptionally substituted heterocyclic ring, e.g., an optionallysubstituted 3-to 6-membered heterocyclic ring. In certain embodiments,one R^(N) group and one R^(D) group are joined to form an optionallysubstituted 5-membered heterocyclic ring, or optionally substituted6-membered heterocyclic ring. In certain embodiments, one R^(N) groupand one R^(D) group are joined to form an unsubstituted heterocyclicring. However, in certain embodiments, one R^(N) group and one R^(D)group are joined to form a substituted heterocyclic ring, e.g.,substituted with one or more alkyl groups.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-18)-(q-31):

wherein V²¹, V²², V²³, V²⁴, and R^(D) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-32)-(q-45):

wherein V²¹, V²², V²³, V²⁴, and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-46)-(q-49):

wherein V²¹, V²², V²³, V²⁴, R^(A), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-50)-(q-57):

wherein V²¹, V²², V²³, V²⁴, and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-58)-(q-72):

wherein V²¹, V²², V²³, V²⁴, R^(A), R^(D), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula (q-73) and(q-76)-(q-79):

wherein V²¹, V²², V²³, V²⁴, R^(A), R^(D), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula (q-74),(q-75) and (q-80)-(q-87):

wherein V²¹, V²², V²³, V²⁴, R^(A), R^(D), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-88)-(q-102):

wherein V²¹, V²², V²³, V²⁴, R^(D), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴⁾ is of Formula(q-103)-(q-117):

wherein V²¹, V²², V²³, V²⁴, R^(D), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-7a)-(q-17a):

wherein R^(D), R^(N), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-18a)-(q-31a):

wherein R^(D) is defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-32a)-(q-45a):

wherein R^(F) is defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-46a)-(q-49a):

wherein R^(A) and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-50a)-(q-57a):

wherein R^(F) is defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-58a)-(q-72a):

wherein R^(A), R^(D), and R^(F) are defined herein.

In certain embodiments, R^(W) (or Ring A) or R⁴ (as provided in theabove recited instance) is of Formula (q-73a), and (q-76a)-(q-79a):

wherein R^(A), R^(D), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula (q-74a),(q-75a) and (q-80a)-(q-87a):

wherein R^(A), R^(D), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-88a)-(q-102a):

wherein R^(D), and R^(F) are defined herein.

In certain embodiments, Ring A (R^(W) or R⁴) is of Formula(q-103a)-(q-117a):

wherein V²¹, V²², V²³, V²⁴, R^(D), and R^(F) are defined herein.

In certain embodiments, wherein, for example, 2 R^(D) groups are joinedto form a C₅₋₆ membered carbocyclic ring, or a 5-6-membered heterocyclicring, Formula (q-7) is of Formula (r-1) and (r-2):

and Formula (q-9) is of Formula (r-3) and (r-4):

wherein:

V²¹, V²², V²³, and V²⁴ are as defined herein;

V²⁶, V²⁷, V²⁸, V²⁹, and V³⁰ are each independently O, S, NR^(Na), C═O,or C(R^(E))₂ as valency permits, provided no more than two of V²⁶, V²⁷,V²⁸, V²⁹, and V³⁰ is a heteroatom O, S, and NR^(Na); alternativelywherein one of V²⁶, V²⁷, V²⁸, V²⁹, and V³⁰ and another of V²⁶, V²⁷, V²⁸,V²⁹, and V³⁰ adjacent to each other are joined to form an N═C(R^(E)) orC(R^(E))═C(R^(E)) group;

each instance of R^(E) is independently hydrogen, halo, —CN, —NO₂,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted carbocyclyl, optionallysubstituted aryl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or two R^(E) groups are joined toform an optionally substituted carbocyclic or optionally substitutedheterocyclic ring; and

each instance of R^(Na) is independently hydrogen, optionallysubstituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, optionallysubstituted heteroaryl, —C(═O)R^(A), —C(═O)OR^(A), —C(═O)SR^(A),—C(═O)N(R^(B))₂, —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —C(═S)R^(A), —C(═S)N(R^(B))₂, —S(═O)R^(A),—SO₂R^(A), —SO₂N(R^(B))₂, or a nitrogen protecting group.

In certain embodiments, Formula (q-7) or (q-9) is of Formula (r-5) to(r-24):

wherein V²¹, V²², V²³, V²⁴, V²⁶, V²⁷, V²⁸, V²⁹, V³⁰, and R^(Na) aredefined herein.

In certain embodiments, Formula (q-7) or (q-9) is of Formula (r-25) to(r-44):

wherein V²¹, V²², V²³, V²⁴, V²⁶, V²⁷, V²⁸, V²⁹, V³⁰, and R^(Na) aredefined herein.

In certain embodiments, Formula (q-7) or (q-9) is of Formula (r-45) to(r-68):

wherein V²¹, V²², V²³, V²⁴, V²⁶, V²⁷, V²⁸, V²⁹, V³⁰, R^(Na), and R^(A)are defined herein.

In certain embodiments, Formula (q-7) or (q-9) is of Formula (r-69) to(r-76):

wherein V²¹, V²², V²³, V²⁴, V²⁶, V²⁷, V²⁸, V²⁹, V³⁰, and R^(Na) aredefined herein.

In certain embodiments, Formula (q-7) or (q-9) is of Formula (r-77) to(r-99):

wherein V²¹, V²², V²³, V²⁴, R^(E), and R^(Na) are defined herein, and zis 0, 1, 2, 3, or 4.

In certain embodiments, Formula (q-7) or (q-9) is of Formula (r-100) to(r-119):

wherein V²¹, V²², V²³, V²⁴, R^(E), and R^(Na) are defined herein, and zis 0, 1, 2, 3, or 4.

In certain embodiments, Formula (q-7) is of Formula (r-120) to (r-143):

wherein V²¹, V²², V²³, V²⁴, R^(A), R^(E), and R^(Na) are defined herein,and z is 0, 1, 2, 3, or 4.

In certain embodiments, Formula (q-7) is of Formula (r-144) to (r-147):

wherein V²¹, V²², V²³, V²⁴, and R^(E) are defined herein, and z is 0, 1,2, 3, or 4.

In certain embodiments, Formula (q-7) is of Formula (r-148) to (r-161):

wherein R^(E) is as defined herein, and z is 0, 1, 2, 3, or 4.

In certain embodiments, Formula (q-7) is of Formula (r-162) to (r-173):

wherein V²¹, V²², V²³, V²⁴, and R^(E) are defined herein.

In certain embodiments, Formula (q-7) is of Formula (r-174) to (r-185):

wherein R^(E) is defined herein.

In certain embodiments, Formula (q-7) is of Formula (r-186) to (r-193):

wherein V²¹, V²², V²³, V²⁴, and R^(E) are defined herein.

In certain embodiments, Formula (q-7) is of Formula (r-194) to (r-213):

wherein V²¹, V²², V²³, V²⁴, and R^(E) are defined herein.

In certain embodiments, Formula (q-7) is of Formula (r-214) to (r-217):

wherein V²¹, V²², V²³, V²⁴, z, and R^(E) are defined herein.

In certain embodiments, Formula (q-7) is of Formula (r-218)-(r-227):

wherein V²¹, V²², V²³, V²⁴, z, and R^(E) are defined herein;

each instance of R^(G) is independently hydrogen, halo, optionallysubstituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, or optionally substituted carbocyclyl; or two R^(G)groups can be taken together to form an optionally substitutedcarbocyclic ring; and

R^(H) is hydrogen, halo, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted aryl, optionallysubstituted heterocyclyl, or optionally substituted heteroaryl.

In certain embodiments, Formula (q-7) is of Formula (r-228) to (r-230):

wherein V²¹, V²², V²³, V²⁴, R^(E), and R^(G) are defined herein;

z is 0, 1, 2, 3, or 4;

each instance of A is independently N or CR^(Ha) provided that no morethan 2 instances of A can be N; and

each instance of R^(Ha) is independently hydrogen, halo, —CN, —NO₂,optionally substituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted carbocyclyl, optionallysubstituted aryl, optionally substituted heterocyclyl, optionallysubstituted heteroaryl, —OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A),—C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂,—OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂,—NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A), —SC(O)R^(A),—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂.

In certain embodiments, Formula (q-7) is of Formula (r-231) to (r-233):

wherein V²¹, V²², V²³, V²⁴, z, R^(E), R^(G), and R^(Ha) are definedherein.

In certain embodiments, Formula (q-7) is of Formula (r-234) to (r-236):

wherein V²¹, V²², V²³, V²⁴, z, R^(E), R^(G), and R^(Ha) are definedherein.

In some embodiments, -L₁-R^(W) is optionally substituted carbocyclyl. Insome embodiments, -L₁-R^(W) is optionally substituted C₃₋₆ carbocyclyl.In some embodiments, -L₁-R^(W) is unsubstituted cyclopropyl. In someembodiments, -L₁-R^(W) is substituted cyclopropyl. In some embodiments,-L₁-R^(W) is unsubstituted cyclobutyl. In some embodiments, -L₁-R^(W) issubstituted cyclobutyl. In some embodiments, -L₁-R^(W) is unsubstitutedcyclopentyl. In some embodiments, -L₁-R^(W) is substituted cyclopentyl.In some embodiments, -L₁-R^(W) is unsubstituted cyclohexyl. In someembodiments, -L₁-R^(W) is substituted cyclohexyl. In some embodiments,-L₁-R^(W) is optionally substituted cyclopentenyl or optionallysubstituted cyclohexenyl.

In some embodiments, -L₁-R^(W) is optionally substituted heterocyclyl.In some embodiments, -L₁-R^(W) is an optionally substituted 4-to7-membered heterocyclyl ring having 1-2 heteroatoms independentlyselected from nitrogen, oxygen, and sulfur. In some embodiments,-L₁-R^(W) is azetidinyl or oxetanyl. In some embodiments, -L₁-R^(W) isoptionally substituted tetrahydrofuranyl, optionally substitutedpyrrolidinyl, optionally substituted dihydropyrrolyl, or optionallysubstituted pyrrolyl-2,5-dione. In some embodiments, -L₁-R^(W) isoptionally substituted piperidinyl, optionally substitutedtetrahydropyranyl, optionally substituted dihydropyranyl, optionallysubstituted dihydropyridinyl, and optionally substituted thianyl. Incertain embodiments, -L₁-R^(W) is optionally substituted piperidinyl. Insome embodiments, -L₁-R^(W) is optionally substituted piperazinyl,optionally substituted morpholinyl, optionally substituted dithianyl,and optionally substituted dioxanyl. In some embodiments, -L₁-R^(W) is a5- or 6-membered heterocyclyl group fused to a C₆ aryl ring. In someembodiments, -L₁-R^(W) is optionally substituted indolinyl, optionallysubstituted isoindolinyl, optionally substituted dihydrobenzofuranyl,optionally substituted dihydrobenzothienyl, or optionally substitutedbenzoxazolinonyl. In some embodiments, -L₁-R^(W) is optionallysubstituted tetrahydroquinolinyl or optionally substitutedtetrahydroisoquinolinyl.

In some embodiments, -L₁-R^(W) is optionally substituted alkyl. In someembodiments, -L₁-R^(W) is optionally substituted alkenyl. In someembodiments, -L₁-R^(W) is optionally substituted alkynyl. In someembodiments, -L₁-R^(W) is optionally substituted C₁₋₆ alkyl. In someembodiments, -L₁-R^(W) is unsubstituted C₁₋₆ alkyl. In some embodiments,-L₁-R^(W) is methyl, ethyl, propyl, or butyl. In some embodiments,-L₁-R^(W) is isopropyl, isobutyl, or isoamyl. In some embodiments,-L₁-R^(W) is optionally substituted C₂₋₆ alkenyl. In some embodiments,-L₁-R^(W) is C₂₋₆ alkynyl.

As defined generally above, R³ is hydrogen, C₁₋₄ alkyl, or C₃₋₄cycloalkyl. In certain embodiments, R³ is hydrogen. In certainembodiments, R³ is C₁₋₄ alkyl. In certain embodiments, R³ is methyl. Incertain embodiments, R³ is ethyl. In certain embodiments, R³ is propylor butyl. In certain embodiments, R³ is cyclopropyl. In certainembodiment, R³ is cyclobutyl.

As defined generally above, R⁴ is hydrogen, optionally substituted C₁₋₆alkyl, optionally substituted C₂₋₆ alkenyl, optionally substituted C₂₋₆alkynyl, optionally substituted C₃₋₇ cycloalkyl, optionally substituted4-to 7-membered heterocyclyl; or optionally substituted C₁₋₄ alkyl-Cy,wherein Cy is optionally substituted C₃₋₇ cycloalkyl, optionallysubstituted 4-to 7-membered heterocyclyl, optionally substituted aryl,or optionally substituted heteroaryl. Alternatively, when L₁ is a bondand R^(W) is hydrogen, then X is CR⁵, Z is N, and Y is NR⁴, then R⁴ isoptionally substituted carbocyclyl or optionally substitutedheterocyclyl as defined above and herein.

In certain embodiments, R⁴ is hydrogen. In certain embodiments, R⁴ isoptionally substituted C₁₋₆ alkyl. In certain embodiments, R⁴ isunsubstituted C₁₋₆ alkyl. In certain embodiments, R⁴ is methyl, ethyl,or isopropyl. In certain embodiments, R⁴ is substituted C₁-6 alkyl. Incertain embodiments, R⁴ is methoxyethyl. In certain embodiments, R⁴ ishydroxyethyl or propane-1,2-diol. In certain embodiments, R⁴ isoptionally substituted C₃₋₇ cycloalkyl. In certain embodiments, R⁴ isunsubstituted C₃₋₇ cycloalkyl. In certain embodiments, R⁴ iscyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl. In certainembodiments, R⁴ is optionally substituted 4-to 7-membered heterocyclyl.In certain embodiments, R⁴ is optionally substituted 4-to 7-memberedheterocyclyl having 1-2 heteroatoms independently selected fromnitrogen, oxygen, and sulfur. In certain embodiments, R⁴ is oxetane,tetrahydrofuran, or tetrahydropyran.

In certain embodiments, R⁴ is optionally substituted C₁₋₄ alkyl-Cy,wherein Cy is optionally substituted C₃₋₇ cycloalkyl, optionallysubstituted 4-to 7-membered heterocyclyl, optionally substituted aryl,or optionally substituted heteroaryl. In some embodiments, Cy isoptionally substituted C₃₋₇ cycloalkyl. In some embodiments, Cy isoptionally substituted 4- to 7-membered heterocyclyl having 1-2heteroatoms independently selected from nitrogen, oxygen, and sulfur. Insome embodiments, Cy is oxetane, tetrahydrofuran, or tetrahydropyran. Insome embodiments, Cy is optionally substituted aryl. In someembodiments, Cy is optionally substituted phenyl. In some embodiments,Cy is unsubstituted phenyl. In some embodiments, Cy is optionallysubstituted heteroaryl having 1-3 heteroatoms independently selectedfrom nitrogen, oxygen, and sulfur. In some embodiments, Cy is optionallysubstituted 5-to 6-membered heteroaryl having 1-3 heteroatomsindependently selected from nitrogen, oxygen, and sulfur. In someembodiments, Cy is pyridyl. In some embodiments, R⁴ is

In some embodiments, R⁴ is

In some embodiments, R⁴ is

As defined generally above, R⁵ is hydrogen, halo, —CN, optionallysubstituted C₁₋₄ alkyl, or optionally substituted C₃₋₄ cycloalkyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, R⁵ is halo.In certain embodiments, R⁵ is chloro. In certain embodiments, R⁵ isfluoro. In certain embodiments, R⁵ is —CN. In certain embodiments, R⁵ isoptionally substituted C₁₋₄ alkyl. In certain embodiments, R⁵ isunsubstituted C₁₋₄ alkyl. In certain embodiments, R⁵ is methyl. Incertain embodiments, R⁵ is ethyl. In certain embodiments, R⁵ is propylor butyl. In certain embodiments, R⁵ is C₁₋₄ alkyl substituted with oneor more fluoro groups. In certain embodiments, R⁵ is —CF₃. In certainembodiments, R⁵ is —CHF₂. In certain embodiments, R⁵ is optionallysubstituted C₃₋₄ cycloalkyl. In certain embodiments, R⁵ is cyclopropylor cyclobutyl.

As defined generally above, R^(x) is optionally substituted C₁₋₄ alkylor optionally substituted C₃₋₄ cycloalkyl. In certain embodiments, R^(x)is optionally substituted C₁₋₄ alkyl. In certain embodiments, R^(x) isunsubstituted C₁₋₄ alkyl. In certain embodiments, R^(x) is methyl. Incertain embodiments, R^(x) is ethyl. In certain embodiments, R^(x) isisopropyl. In certain embodiments, R^(x) is propyl or butyl. In certainembodiments, R^(x) is substituted C₁₋₄ alkyl. In certain embodiments,R^(x) is C₁₋₄ alkyl substituted with hydroxyl or alkoxy. In certainembodiments, R^(x) is hydroxyethyl or methoxyethyl. In certainembodiments, R^(x) is optionally substituted C₃₋₄ cycloalkyl. In certainembodiments, R^(x) is unsubstituted C₃₋₄ cycloalkyl. In certainembodiments, R^(x) is cyclopropyl. In certain embodiments, R^(x) iscyclobutyl.

As defined generally above, each R^(A) is independently selected fromthe group consisting of hydrogen, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl, anoxygen protecting group when attached to an oxygen atom, and a sulfurprotecting group when attached to a sulfur atom. In some embodiments,R^(A) is hydrogen. In some embodiments, R^(A) is optionally substitutedalkyl. In some embodiments, R^(A) is optionally substituted alkylsubstituted with a Cy group to form optionally substituted alkyl-Cy,wherein Cy is described herein. In some embodiments, R^(A) is optionallysubstituted alkenyl or optionally substituted alkynyl. In someembodiments, R^(A) is optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, or optionallysubstituted heteroaryl. In some embodiments, R^(A) is an oxygenprotecting group when attached to an oxygen atom. In some embodiments,R^(A) is not an oxygen protecting group. In some embodiments, R^(A) issulfur protecting group when attached to an sulfur atom. In someembodiments, R^(A) is not a sulfur protecting group.

As defined generally above, each R^(B) is independently selected fromthe group consisting of hydrogen, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, and optionally substituted heteroaryl, anda nitrogen protecting group, or two R^(B) groups or an R^(B) group andan R^(W) group on the same nitrogen are taken together with theirintervening atoms to form an optionally substituted heterocyclic ring.In some embodiments, R^(B) is hydrogen. In some embodiments, R^(B) isoptionally substituted alkyl. In some embodiments, R^(B) is optionallyalkyl substituted with a Cy group to form optionally substitutedalkyl-Cy, wherein Cy is described herein. In some embodiments, R^(B) isoptionally substituted alkenyl or optionally substituted alkynyl. Insome embodiments, R^(B) is optionally substituted carbocyclyl,optionally substituted heterocyclyl, optionally substituted aryl, oroptionally substituted heteroaryl. In some embodiments, R^(B) is anitrogen protecting group. In some embodiments, R^(B) is not a nitrogenprotecting group. In some embodiments, two R^(B) groups are takentogether with their intervening atoms to form an optionally substitutedheterocyclic ring. In some embodiments, an R^(B) group and an R^(W)group on the same nitrogen are taken together with their interveningatoms to form an optionally substituted heterocyclic ring.

As defined generally above, each instance of R^(E) is independentlyhydrogen, halo, —CN, —NO₂, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted aryl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or two R^(E)groups are joined to form an optionally substituted carbocyclic oroptionally substituted heterocyclic ring. In certain embodiments, R^(E)is hydrogen. In certain embodiments, R^(E) is not hydrogen. In certainembodiments, R^(E) is halo (e.g., fluoro, chloro, bromo, or iodo). Incertain embodiments, R^(E) is optionally substituted alkyl. In certainembodiments, R^(E) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R^(E) is unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl,propyl, butyl, pentyl, or hexyl). In certain R^(E) is unsubstitutedbranched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl, tert-butyl, isopentyl,neopentyl, or 3-pentyl). In certain embodiments, R^(E) is methyl. Incertain embodiments, R^(E) is C₁₋₆ haloalkyl (e.g., —CF₃, —CF₂H, or—CF₂CH₃). In certain embodiments, R^(E) is —CF₃. In certain embodiments,R^(E) is alkoxyalkyl (e.g. —CH₂OR^(A) or —CH₂CH₂OR^(A)). In certainembodiments, R^(E) is an optionally substituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclopropyl). In certainembodiments, R^(E) is an unsubstituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclopropyl). In certainembodiments, R^(E) is substituted phenyl. In certain embodiments, R^(E)is unsubstituted phenyl. In certain embodiments, R^(E) is an optionallysubstituted heterocyclic ring (e.g., azetidine, oxetane, furan,pyrrolidine, piperidine, piperazine, or morpholine). In certainembodiments, R^(E) is an unsubstituted heterocyclic ring (e.g.,azetidine, oxetane, furan, pyrrolidine, piperidine, piperazine, ormorpholine). In certain embodiments, R^(E) is an optionally substitutedheteroaryl ring (e.g., pyrazole, imidazole, triazole, pyridine,pyrimidine, or pyridizine). In certain embodiments, R^(E) is anunsubstituted heteroaryl ring (e.g., pyrazole, imidazole, triazole,pyridine, pyrimidine, or pyridizine). In certain embodiments, R^(E) isalkoxy. In certain embodiments, R^(E) is —OR^(A); and R^(A) isoptionally substituted C₁₋₆ alkyl. In certain embodiments, R^(E) is—OR^(A); and R^(A) is unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, orpropyl). In certain embodiments, two R^(E) groups are joined to form anoptionally substituted carbocyclic or optionally substitutedheterocyclic ring. In certain embodiments, two R^(E) groups are joinedto form an optionally substituted carbocyclic ring (e.g., cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl). In certain embodiments, twoR^(E) groups are joined to form an optionally substituted cyclopropylring. In certain embodiments, two R^(E) groups are joined to form anunsubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an unsubstituted cyclopropyl ring.

Various combinations of certain above-described embodiments are furtherenvisioned herein.

For example, in certain embodiments, provided are compounds of FormulaeXII-a1 to XII-a5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, andR^(x) are defined herein; and Ring A is any of Formulae (q-7) to(q-117), (q-7a) to (q-117a), or (r-1) to (r-236). In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, L₁ is abond and R^(W) is hydrogen. In certain embodiments, L₁ is a bond, R^(W)is hydrogen, and R⁵ is hydrogen. In certain embodiments, L₁ is a bondand R^(W) is halogen, e.g., fluoro, chloro, bromo, or iodo. In certainembodiments, L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl,e.g., unsubstituted C₁₋₄alkyl, e.g., methyl, ethyl, propyl, butyl,isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (q-7), provided arecompounds of Formulae XII-b1 to XII-b5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, and R^(D) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₂ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₂ is O. In certain embodiments, V₂₂ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, at least one instance of R^(D) is hydrogen. Incertain embodiments, at least one instance of R^(D) is halo (e.g.,fluoro, chloro, or bromo). In certain embodiments, at least one instanceof R^(D) is optionally substituted alkyl. In certain embodiments, atleast one instance of R^(D) is optionally substituted C₁₋₄ alkyl. Incertain embodiments, at least one instance of R^(D) is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, at least one instance of R^(D) is unsubstituted branchedC₃₋₄ alkyl (e.g., isopropyl, isobutyl, sec-butyl, or tert-butyl). Incertain embodiments, one instance of R^(D) is hydrogen; and the secondinstance of R^(D) is optionally substituted alkyl. In certainembodiments, one instance of R^(D) is hydrogen; and the second instanceof R^(D) is optionally substituted C₁₋₄ alkyl. In certain embodiments,one instance of R^(D) is hydrogen; and the second instance of R^(D) isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, one instance of R^(D) is hydrogen; and the secondinstance of R^(D) is branched C₃₋₄ alkyl (e.g., isopropyl, isobutyl,sec-butyl, or tert-butyl). In certain embodiments, one instance of R^(D)is hydrogen; and the second instance of R^(D) is methyl. In certainembodiments, one instance of R^(D) is hydrogen; and the second instanceof R^(D) is ethyl. In certain embodiments, one instance of R^(D) ishydrogen; and the second instance of R^(D) is isopropyl. In certainembodiments, one instance of R^(D) is hydrogen; and the second instanceof R^(D) is isobutyl. In certain embodiments, both instances of R^(D)are optionally substituted C₁₋₄ alkyl. In certain embodiments, bothinstances of R^(D) are methyl. In certain embodiments, one instance ofR^(D) is hydrogen; and one instance of R^(D) is —OR^(A). In certainembodiments, at least one instance of R^(D) is optionally substitutedalkoxyalkyl (e.g., —CH₂OR^(A), —CH₂CH₂OR^(A), or —CH₂CH₂CH₂OR^(A)). Incertain embodiments, both instances of R^(D) is optionally substitutedalkoxyalkyl (e.g., —CH₂OR^(A), —CH₂CH₂OR^(A), or —CH₂CH₂CH₂OR^(A)). Incertain embodiments, two R^(D) groups are joined to form an optionallysubstituted carbocyclic or optionally substituted heterocyclic ring. Incertain embodiments, two R^(D) groups are joined to form an optionallysubstituted cyclopentane. In certain embodiments, two R^(D) groups arejoined to form an optionally substituted cyclohexane. In certainembodiments, two R^(D) groups are joined to form an optionallysubstituted furan. In certain embodiments, two R^(D) groups are joinedto form an optionally substituted pyran. In certain embodiments, twoR^(D) groups are joined to form an optionally substituted pyrrolidinone.In certain embodiments, two R^(E) groups are joined to form anoptionally substituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an optionally substituted cyclopropyl ring. Incertain embodiments, two R^(E) groups are joined to form anunsubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an unsubstituted cyclopropyl ring. In certainembodiments, L₁ is a bond and R^(W) is hydrogen. In certain embodiments,L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. In certainembodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro, chloro,bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W) isoptionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl, e.g.,methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (q-9), provided arecompounds of Formulae XII-c1 to XII-c5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, and R^(D) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₁ and V₂₂ is O or —CH(OR^(A))—. In certain embodiments,V₂₂ is O. In certain embodiments, V₂₂ is —CH(OR^(A))—. In certainembodiments, V₂₁ is O. In certain embodiments, V₂₁ is —CH(OR^(A))—. Incertain embodiments, at least one instance of R^(D) is hydrogen. Incertain embodiments, at least one instance of R^(D) is halo (e.g.,fluoro, chloro, or bromo). In certain embodiments, at least one instanceof R^(D) is optionally substituted alkyl. In certain embodiments, atleast one instance of R^(D) is optionally substituted C₁₋₄ alkyl. Incertain embodiments, at least one instance of R^(D) is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, at least one instance of R^(D) is unsubstituted branchedC₃₋₄ alkyl (e.g., isopropyl, isobutyl, sec-butyl, or tert-butyl). Incertain embodiments, one instance of R^(D) is hydrogen; and the secondinstance of R^(D) is optionally substituted alkyl. In certainembodiments, one instance of R^(D) is hydrogen; and the second instanceof R^(D) is optionally substituted C₁₋₄ alkyl. In certain embodiments,one instance of R^(D) is hydrogen; and the second instance of R^(D) isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, one instance of R^(D) is hydrogen; and the secondinstance of R^(D) is branched C₃₋₄ alkyl (e.g., isopropyl, isobutyl,sec-butyl, or tert-butyl). In certain embodiments, one instance of R^(D)is hydrogen; and the second instance of R^(D) is methyl. In certainembodiments, one instance of R^(D) is hydrogen; and the second instanceof R^(D) is ethyl. In certain embodiments, one instance of R^(D) ishydrogen; and the second instance of R^(D) is isopropyl. In certainembodiments, one instance of R^(D) is hydrogen; and the second instanceof R^(D) is isobutyl. In certain embodiments, both instances of R^(D)are optionally substituted C₁₋₄ alkyl. In certain embodiments, bothinstances of R^(D) are methyl. In certain embodiments, one instance ofR^(D) is hydrogen; and one instance of R^(D) is —OR^(A). In certainembodiments, at least one instance of R^(D) is optionally substitutedalkoxyalkyl (e.g., —CH₂OR^(A), —CH₂CH₂OR^(A), or —CH₂CH₂CH₂OR^(A)). Incertain embodiments, both instances of R^(D) is optionally substitutedalkoxyalkyl (e.g., —CH₂OR^(A), —CH₂CH₂OR^(A), or —CH₂CH₂CH₂OR^(A)). Incertain embodiments, two R^(D) groups are joined to form an optionallysubstituted carbocyclic or optionally substituted heterocyclic ring. Incertain embodiments, two R^(D) groups are joined to form an optionallysubstituted cyclopentane. In certain embodiments, two R^(D) groups arejoined to form an optionally substituted cyclohexane. In certainembodiments, two R^(D) groups are joined to form an optionallysubstituted furan. In certain embodiments, two R^(D) groups are joinedto form an optionally substituted pyran. In certain embodiments, twoR^(D) groups are joined to form an optionally substituted pyrrolidinone.In certain embodiments, two R^(E) groups are joined to form anoptionally substituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an optionally substituted cyclopropyl ring. Incertain embodiments, two R^(E) groups are joined to form anunsubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an unsubstituted cyclopropyl ring. In certainembodiments, L₁ is a bond and R^(W) is hydrogen. In certain embodiments,L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. In certainembodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro, chloro,bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W) isoptionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl, e.g.,methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (q-46) or (q-47),provided are compounds of Formulae XII-d1 to XII-d5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, R^(A), and R^(F) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₂ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₂ is O. In certain embodiments, V₂₂ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, at least two instances of R^(F) are hydrogen. Incertain embodiments, all four instances of R^(F) are hydrogen. Incertain embodiments, each instance of R^(A) is independently hydrogen.In certain embodiments, both instances of R^(A) are hydrogen. In certainembodiments, neither instance of R^(A) is hydrogen. In certainembodiments, at least one instance of R^(A) is optionally substitutedalkyl. In certain embodiments, both instances of R^(A) are independentlyoptionally substituted alkyl. In certain embodiments, at least oneinstance of R^(A) is optionally substituted C₁₋₄ alkyl. In certainembodiments, both instances of R^(A) are independently optionallysubstituted C₁₋₄ alkyl. In certain embodiments, at least one instance ofR^(A) is unsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, orbutyl). In certain embodiments, both instances of R^(A) areindependently unsubstituted alkyl (e.g., methyl, ethyl, propyl, orbutyl). In certain embodiments, at least one instance of R^(A) ismethyl. In certain embodiments, both instances of R^(A) are methyl. Incertain embodiments, at least one instance of R^(A) is ethyl. In certainembodiments, both instances of R^(A) are ethyl. In certain embodiments,two R^(E) groups are joined to form an optionally substitutedcarbocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl). In certain embodiments, two R^(E) groups are joined to forman optionally substituted cyclopropyl ring. In certain embodiments, twoR^(E) groups are joined to form an unsubstituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an unsubstitutedcyclopropyl ring. In certain embodiments, L₁ is a bond and R^(W) ishydrogen. In certain embodiments, L₁ is a bond, R^(W) is hydrogen, andR⁵ is hydrogen. In certain embodiments, L₁ is a bond and R^(W) ishalogen, e.g., fluoro, chloro, bromo, or iodo. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl, e.g.,unsubstituted C₁₋₄alkyl, e.g., methyl, ethyl, propyl, butyl, isopropyl,isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (q-58), provided arecompounds of Formulae XII-e1 to XII-e5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, R^(A), and R^(D) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₂ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₂ is O. In certain embodiments, V₂₂ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, R^(D) is hydrogen. In certain embodiments, R^(A) ishydrogen. In certain embodiments, R^(A) is not hydrogen. In certainembodiments, R^(A) is optionally substituted alkyl. In certainembodiments, R^(A) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R^(A) is unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl,propyl, butyl, pentyl, or hexyl). In certain embodiments, R^(A) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,R^(A) is methyl. In certain embodiments, R^(A) is ethyl. In certainembodiments, R^(A) is 3-pentyl. In certain embodiments, R^(D) ishydrogen; and R^(A) is methyl. In certain embodiments, R^(D) ishydrogen; and R^(A) is ethyl. In certain embodiments, R^(D) is hydrogen;and R^(A) is 3-pentyl. In certain embodiments, two R^(E) groups arejoined to form an optionally substituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an optionallysubstituted cyclopropyl ring. In certain embodiments, two R^(E) groupsare joined to form an unsubstituted carbocyclic ring (e.g., cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl). In certain embodiments, twoR^(E) groups are joined to form an unsubstituted cyclopropyl ring. Incertain embodiments, L₁ is a bond and R^(W) is hydrogen. In certainembodiments, L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. Incertain embodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro,chloro, bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W)is optionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl,e.g., methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (q-59), provided arecompounds of Formulae XII-f1 to XII-f5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, R^(A), R^(D), and R^(F) are defined herein. Incertain embodiments, R³ is hydrogen. In certain embodiments, R³ isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, R³ is methyl. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g.,methyl, ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl.In certain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₂ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₂ is O. In certain embodiments, V₂₂ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, both instances of R^(F) are hydrogen. In certainembodiments, R^(D) is hydrogen. In certain embodiments, R^(A) ishydrogen. In certain embodiments, R^(A) is not hydrogen. In certainembodiments, R^(A) is optionally substituted alkyl. In certainembodiments, R^(A) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R^(A) is unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl,propyl, butyl, pentyl, or hexyl). In certain embodiments, R^(A) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,R^(A) is methyl. In certain embodiments, R^(A) is ethyl. In certainembodiments, R^(A) is 3-pentyl. In certain embodiments, R^(D) ishydrogen; and R^(A) is methyl. In certain embodiments, R^(D) ishydrogen; and R^(A) is ethyl. In certain embodiments, R^(D) is hydrogen;and

R^(A) is 3-pentyl. In certain embodiments, two R^(E) groups are joinedto form an optionally substituted carbocyclic ring (e.g., cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl). In certain embodiments, twoR^(E) groups are joined to form an optionally substituted cyclopropylring. In certain embodiments, two R^(E) groups are joined to form anunsubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an unsubstituted cyclopropyl ring. In certainembodiments, L₁ is a bond and R^(W) is hydrogen. In certain embodiments,L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. In certainembodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro, chloro,bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W) isoptionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl, e.g.,methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (q-60), provided arecompounds of Formulae XII-g1 to XII-g5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, R^(A), R^(D), and R^(F) are defined herein. Incertain embodiments, R³ is hydrogen. In certain embodiments, R³ isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, R³ is methyl. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g.,methyl, ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl.In certain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₂ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₂ is O. In certain embodiments, V₂₂ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, at least two instances of R^(F) are hydrogen. Incertain embodiments, all four instances of R^(F) are hydrogen. Incertain embodiments, R^(D) is hydrogen. In certain embodiments, R^(A) ishydrogen. In certain embodiments, R^(A) is not hydrogen. In certainembodiments, R^(A) is optionally substituted alkyl. In certainembodiments, R^(A) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R^(A) is unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl,propyl, butyl, pentyl, or hexyl). In certain embodiments, R^(A) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,R^(A) is methyl. In certain embodiments, R^(A) is ethyl. In certainembodiments, R^(A) is 3-pentyl. In certain embodiments, R^(D) ishydrogen; and R^(A) is methyl. In certain embodiments, R^(D) ishydrogen; and R^(A) is ethyl. In certain embodiments, R^(D) is hydrogen;and R^(A) is 3-pentyl. In certain embodiments, two R^(E) groups arejoined to form an optionally substituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an optionallysubstituted cyclopropyl ring. In certain embodiments, two R^(E) groupsare joined to form an unsubstituted carbocyclic ring (e.g., cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl). In certain embodiments, twoR^(E) groups are joined to form an unsubstituted cyclopropyl ring. Incertain embodiments, L₁ is a bond and R^(W) is hydrogen. In certainembodiments, L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. Incertain embodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro,chloro, bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W)is optionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl,e.g., methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-1), provided arecompounds of Formulae XII-h1 to XII-h5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, V₂₆, V₂₇, V₂₈, and V₂₉ are defined herein. Incertain embodiments, R³ is hydrogen. In certain embodiments, R³ isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, R³ is methyl. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g.,methyl, ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl.In certain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₂ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₂ is O. In certain embodiments, V₂₂ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈, or V₂₉ is O.In certain embodiments, exactly two instances of V₂₆, V₂₇, V₂₈, or V₂₉are O. In certain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈, orV₂₉ is O; and all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certainembodiments, exactly one instance of V₂₆, V₂₇, V₂₈, or V₂₉ is O; and asecond instance of V₂₆, V₂₇, V₂₈, or V₂₉ is —C(R^(E))₂—. In certainembodiments, exactly one instance of V₂₆, V₂₇, V₂₈, or V₂₉ is O; and asecond instance of V₂₆, V₂₇, V₂₈, or V₂₉ is —C(Me)₂-. In certainembodiments, all four of V₂₆, V₂₇, V₂₈, and V₂₉ are —CH₂—. In certainembodiments, all eight of V₂₁, V₂₂, V₂₃, V₂₄, V₂₆, V₂₇, V₂₈, and V₂₉ are—CH₂—. In certain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈, orV₂₉ is NR^(Na); and a second instance of V₂₆, V₂₇, V₂₈, or V₂₉ is C═O.In certain embodiments, at least one instance of V₂₆, V₂₇, V₂₈, or V₂₉is —C(R^(E))₂—. In certain embodiments, exactly one instance of V₂₆,V₂₇, V₂₈, or V₂₉ is —C(R^(E))₂—; and each instance of R^(E) isindependently halogen (e.g., fluoro, chloro, or bromo). In certainembodiments, two R^(E) groups are joined to form an optionallysubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an optionally substituted cyclopropyl ring. Incertain embodiments, two R^(E) groups are joined to form anunsubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an unsubstituted cyclopropyl ring. In certainembodiments, L₁ is a bond and R^(W) is hydrogen. In certain embodiments,L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. In certainembodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro, chloro,bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W) isoptionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl, e.g.,methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-3), provided arecompounds of Formulae XII-i1 to XII-i5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, V₂₆, V₂₇, V₂₈, and V₂₉ are defined herein. Incertain embodiments, R³ is hydrogen. In certain embodiments, R³ isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, R³ is methyl. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g.,methyl, ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl.In certain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈, or V₂₉ is O.In certain embodiments, exactly two instances of V₂₆, V₂₇, V₂₈, or V₂₉are O. In certain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈, orV₂₉ is O; and all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certainembodiments, exactly one instance of V₂₆, V₂₇, V₂₈, or V₂₉ is O; and asecond instance of V₂₆, V₂₇, V₂₈, or V₂₉ is —C(R^(E))₂—. In certainembodiments, exactly one instance of V₂₆, V₂₇, V₂₈, or V₂₉ is O; and asecond instance of V₂₆, V₂₇, V₂₈, or V₂₉ is —C(Me)₂-. In certainembodiments, all four of V₂₆, V₂₇, V₂₈, and V₂₉ are —CH₂—. In certainembodiments, all eight of V₂₁, V₂₂, V₂₃, V₂₄, V₂₆, V₂₇, V₂₈, and V₂₉ are—CH₂—. In certain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈, orV₂₉ is NR^(Na); and a second instance of V₂₆, V₂₇, V₂₈, or V₂₉ is C═O.In certain embodiments, at least one instance of V₂₆, V₂₇, V₂₈, or V₂₉is —C(R^(E))₂—. In certain embodiments, exactly one instance of V₂₆,V₂₇, V₂₈, or V₂₉ is —C(R^(E))₂—; and each instance of R^(E) isindependently halogen (e.g., fluoro, chloro, or bromo). In certainembodiments, two R^(E) groups are joined to form an optionallysubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an optionally substituted cyclopropyl ring. Incertain embodiments, two R^(E) groups are joined to form anunsubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an unsubstituted cyclopropyl ring. In certainembodiments, L₁ is a bond and R^(W) is hydrogen. In certain embodiments,L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. In certainembodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro, chloro,bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W) isoptionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl, e.g.,methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-2), provided arecompounds of Formulae XII-j1 to XII-j5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, V₂₆, V₂₇, V₂₈, V₂₉, and V₃₀ are defined herein. Incertain embodiments, R³ is hydrogen. In certain embodiments, R³ isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, R³ is methyl. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g.,methyl, ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl.In certain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₂ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₂ is O. In certain embodiments, V₂₂ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀is O. In certain embodiments, exactly two instances of V₂₆, V₂₇, V₂₈,V₂₉, or V₃₀ are O. In certain embodiments, exactly one instance of V₂₆,V₂₇, V₂₈, V₂₉, or V₃₀ is O; and all four of V₂₁, V₂₂, V₂₃, and V₂₄ are—CH₂—. In certain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈,V₂₉, or V₃₀ is O; and a second instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀ is—C(R^(E))₂—. In certain embodiments, exactly one instance of V₂₆, V₂₇,V₂₈, V₂₉, or V₃₀ is O; and a second instance of V₂₆, V₂₇, V₂₈, V₂₉, orV₃₀ is —C(Me)₂-. In certain embodiments, all five of V₂₆, V₂₇, V₂₈, V₂₉,and V₃₀ are —CH₂—. In certain embodiments, all nine of V₂₁, V₂₂, V₂₃,V₂₄, V₂₆, V₂₇, V₂₈, V₂₉, and V₃₀ are —CH₂—. In certain embodiments,exactly one instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀ is NR^(Na); and asecond instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀ is C═O. In certainembodiments, at least one instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀ is—C(R^(E))₂—. In certain embodiments, exactly one instance of V₂₆, V₂₇,V₂₈, V₂₉, or V₃₀ is —C(R^(E))₂—; and each instance of R^(E) isindependently halogen (e.g., fluoro, chloro, or bromo). In certainembodiments, V₃₀ is —CF₂—; and V₂₆, V₂₇, V₂₈, and V₂₉ are —CH₂—. Incertain embodiments, V₃₀ is —CF₂—; and V₂₁, V₂₂, V₂₃, V₂₄, V₂₆, V₂₇,V₂₈, and V₂₉ are —CH₂—. In certain embodiments, two R^(E) groups arejoined to form an optionally substituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an optionallysubstituted cyclopropyl ring. In certain embodiments, two R^(E) groupsare joined to form an unsubstituted carbocyclic ring (e.g., cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl). In certain embodiments, twoR^(E) groups are joined to form an unsubstituted cyclopropyl ring. Incertain embodiments, L₁ is a bond and R^(W) is hydrogen. In certainembodiments, L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. Incertain embodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro,chloro, bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W)is optionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl,e.g., methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-4), provided arecompounds of Formulae XII-k1 to XII-k5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, V₂₆, V₂₇, V₂₈, V₂₉, and V₃₀ are defined herein. Incertain embodiments, R³ is hydrogen. In certain embodiments, R³ isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, R³ is methyl. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g.,methyl, ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl.In certain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀is O. In certain embodiments, exactly two instances of V₂₆, V₂₇, V₂₈,V₂₉, or V₃₀ are O. In certain embodiments, exactly one instance of V₂₆,V₂₇, V₂₈, V₂₉, or V₃₀ is O; and all four of V₂₁, V₂₂, V₂₃, and V₂₄ are—CH₂—. In certain embodiments, exactly one instance of V₂₆, V₂₇, V₂₈,V₂₉, or V₃₀ is O; and a second instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀ is—C(R^(E))₂—. In certain embodiments, exactly one instance of V₂₆, V₂₇,V₂₈, V₂₉, or V₃₀ is O; and a second instance of V₂₆, V₂₇, V₂₈, V₂₉, orV₃₀ is —C(Me)₂-. In certain embodiments, all five of V₂₆, V₂₇, V₂₈, V₂₉,and V₃₀ are —CH₂—. In certain embodiments, all nine of V₂₁, V₂₂, V₂₃,V₂₄, V₂₆, V₂₇, V₂₈, V₂₉, and V₃₀ are —CH₂—. In certain embodiments,exactly one instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀ is NR^(Na); and asecond instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀ is C═O. In certainembodiments, at least one instance of V₂₆, V₂₇, V₂₈, V₂₉, or V₃₀ is—C(R^(E))₂—. In certain embodiments, exactly one instance of V₂₆, V₂₇,V₂₈, V₂₉, or V₃₀ is —C(R^(E))₂—; and each instance of R^(E) isindependently halogen (e.g., fluoro, chloro, or bromo). In certainembodiments, V₃₀ is —CF₂—; and V₂₆, V₂₇, V₂₈, and V₂₉ are —CH₂—. Incertain embodiments, V₃₀ is —CF₂—; and V₂₁, V₂₂, V₂₃, V₂₄, V₂₆, V₂₇,V₂₈, and V₂₉ are —CH₂—. In certain embodiments, two R^(E) groups arejoined to form an optionally substituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an optionallysubstituted cyclopropyl ring. In certain embodiments, two R^(E) groupsare joined to form an unsubstituted carbocyclic ring (e.g., cyclopropyl,cyclobutyl, cyclopentyl, or cyclohexyl). In certain embodiments, twoR^(E) groups are joined to form an unsubstituted cyclopropyl ring. Incertain embodiments, L₁ is a bond and R^(W) is hydrogen. In certainembodiments, L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. Incertain embodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro,chloro, bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W)is optionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl,e.g., methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-81), provided arecompounds of Formulae XII-I1 to XII-15:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, z and R^(E) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, z is 2. In certain embodiments, at least oneinstance of R^(E) is optionally substituted alkyl. In certainembodiments, at least one instance of R^(E) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, at least one instance of R^(E) isunsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, orhexyl). In certain embodiments, at least one instance of R^(E) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least one instance of R^(E) is methyl. In certain embodiments, atleast two instances of R^(E) are independently optionally substitutedalkyl. In certain embodiments, at least two instances of R^(E) areindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,at least two instances of R^(E) are independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, at least two instances of R^(E) are independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least two instances of R^(E) are methyl. In certain embodiments, z is2; and each instance of R^(E) is independently optionally substitutedalkyl. In certain embodiments, z is 2; and each instance of R^(E) isindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,z is 2; and each instance of R^(E) is independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, z is 2; and each instance of R^(E) is independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,z is 2; and each instance of R^(E) is methyl. In certain embodiments, zis 2; both instances of R^(E) are attached to the same carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; both instances of R^(E) are attached to thesame carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; both instances of R^(E) are attached to the samecarbon; and each instance of R^(E) is methyl. In certain embodiments,two R^(E) groups are joined to form an optionally substitutedcarbocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl). In certain embodiments, two R^(E) groups are joined to forman optionally substituted cyclopropyl ring. In certain embodiments, twoR^(E) groups are joined to form an unsubstituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an unsubstitutedcyclopropyl ring. In certain embodiments, L₁ is a bond and R^(W) ishydrogen. In certain embodiments, L₁ is a bond, R^(W) is hydrogen, andR⁵ is hydrogen. In certain embodiments, L₁ is a bond and R^(W) ishalogen, e.g., fluoro, chloro, bromo, or iodo. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl, e.g.,unsubstituted C₁₋₄alkyl, e.g., methyl, ethyl, propyl, butyl, isopropyl,isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-82), provided arecompounds of Formulae XII-ml to XII-m5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, z and R^(E) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, z is 2. In certain embodiments, at least oneinstance of R^(E) is optionally substituted alkyl. In certainembodiments, at least one instance of R^(E) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, at least one instance of R^(E) isunsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, orhexyl). In certain embodiments, at least one instance of R^(E) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least one instance of R^(E) is methyl. In certain embodiments, atleast two instances of R^(E) are independently optionally substitutedalkyl. In certain embodiments, at least two instances of R^(E) areindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,at least two instances of R^(E) are independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, at least two instances of R^(E) are independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least two instances of R^(E) are methyl. In certain embodiments, z is2; and each instance of R^(E) is independently optionally substitutedalkyl. In certain embodiments, z is 2; and each instance of R^(E) isindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,z is 2; and each instance of R^(E) is independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, z is 2; and each instance of R^(E) is independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,z is 2; and each instance of R^(E) is methyl. In certain embodiments, zis 2; both instances of R^(E) are attached to the same carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; both instances of R^(E) are attached to thesame carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; both instances of R^(E) are attached to the samecarbon; and each instance of R^(E) is methyl. In certain embodiments,two R^(E) groups are joined to form an optionally substitutedcarbocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl). In certain embodiments, two R^(E) groups are joined to forman optionally substituted cyclopropyl ring. In certain embodiments, twoR^(E) groups are joined to form an unsubstituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an unsubstitutedcyclopropyl ring. In certain embodiments, L₁ is a bond and R^(W) ishydrogen. In certain embodiments, L₁ is a bond, R^(W) is hydrogen, andR⁵ is hydrogen. In certain embodiments, L₁ is a bond and R^(W) ishalogen, e.g., fluoro, chloro, bromo, or iodo. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl, e.g.,unsubstituted C₁₋₄alkyl, e.g., methyl, ethyl, propyl, butyl, isopropyl,isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-85), provided arecompounds of Formulae XII-n1 to XII-n5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, z and R^(E) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, z is 2. In certain embodiments, at least oneinstance of R^(E) is optionally substituted alkyl. In certainembodiments, at least one instance of R^(E) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, at least one instance of R^(E) isunsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, orhexyl). In certain embodiments, at least one instance of R^(E) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least one instance of R^(E) is methyl. In certain embodiments, atleast two instances of R^(E) are independently optionally substitutedalkyl. In certain embodiments, at least two instances of R^(E) areindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,at least two instances of R^(E) are independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, at least two instances of R^(E) are independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least two instances of R^(E) are methyl. In certain embodiments, z is2; and each instance of R^(E) is independently optionally substitutedalkyl. In certain embodiments, z is 2; and each instance of R^(E) isindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,z is 2; and each instance of R^(E) is independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, z is 2; and each instance of R^(E) is independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,z is 2; and each instance of R^(E) is methyl. In certain embodiments, zis 2; both instances of R^(E) are attached to the same carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; both instances of R^(E) are attached to thesame carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; both instances of R^(E) are attached to the samecarbon; and each instance of R^(E) is methyl. In certain embodiments, zis 2; each instance of R^(E) is attached to a different carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; each instance of R^(E) is attached to adifferent carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; each instance ofR^(E) is attached to a different carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; each instanceof R^(E) is attached to a different carbon; and each instance of R^(E)is independently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; each instance of R^(E) is attached to a differentcarbon; and each instance of R^(E) is methyl. In certain embodiments,two R^(E) groups are joined to form an optionally substitutedcarbocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl). In certain embodiments, two R^(E) groups are joined to forman optionally substituted cyclopropyl ring. In certain embodiments, twoR^(E) groups are joined to form an unsubstituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an unsubstitutedcyclopropyl ring. In certain embodiments, L₁ is a bond and R^(W) ishydrogen. In certain embodiments, L₁ is a bond, R^(W) is hydrogen, andR⁵ is hydrogen. In certain embodiments, L₁ is a bond and R^(W) ishalogen, e.g., fluoro, chloro, bromo, or iodo. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl, e.g.,unsubstituted C₁₋₄alkyl, e.g., methyl, ethyl, propyl, butyl, isopropyl,isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-86), provided arecompounds of Formulae XII-ol to XII-o5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, z and R^(E) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, z is 2. In certain embodiments, at least oneinstance of R^(E) is optionally substituted alkyl. In certainembodiments, at least one instance of R^(E) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, at least one instance of R^(E) isunsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, orhexyl). In certain embodiments, at least one instance of R^(E) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least one instance of R^(E) is methyl. In certain embodiments, atleast two instances of R^(E) are independently optionally substitutedalkyl. In certain embodiments, at least two instances of R^(E) areindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,at least two instances of R^(E) are independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, at least two instances of R^(E) are independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least two instances of R^(E) are methyl. In certain embodiments, z is2; and each instance of R^(E) is independently optionally substitutedalkyl. In certain embodiments, z is 2; and each instance of R^(E) isindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,z is 2; and each instance of R^(E) is independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, z is 2; and each instance of R^(E) is independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,z is 2; and each instance of R^(E) is methyl. In certain embodiments, zis 2; both instances of R^(E) are attached to the same carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; both instances of R^(E) are attached to thesame carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; both instances of R^(E) are attached to the samecarbon; and each instance of R^(E) is methyl. In certain embodiments, zis 2; each instance of R^(E) is attached to a different carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; each instance of R^(E) is attached to adifferent carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; each instance ofR^(E) is attached to a different carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; each instanceof R^(E) is attached to a different carbon; and each instance of R^(E)is independently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; each instance of R^(E) is attached to a differentcarbon; and each instance of R^(E) is methyl. In certain embodiments,two R^(E) groups are joined to form an optionally substitutedcarbocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl). In certain embodiments, two R^(E) groups are joined to forman optionally substituted cyclopropyl ring. In certain embodiments, twoR^(E) groups are joined to form an unsubstituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an unsubstitutedcyclopropyl ring. In certain embodiments, L₁ is a bond and R^(W) ishydrogen. In certain embodiments, L₁ is a bond, R^(W) is hydrogen, andR⁵ is hydrogen. In certain embodiments, L₁ is a bond and R^(W) ishalogen, e.g., fluoro, chloro, bromo, or iodo. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl, e.g.,unsubstituted C₁₋₄alkyl, e.g., methyl, ethyl, propyl, butyl, isopropyl,isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-87), provided arecompounds of Formulae XII-p1 to XII-p5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, z and R^(E) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, z is 2. In certain embodiments, at least oneinstance of R^(E) is optionally substituted alkyl. In certainembodiments, at least one instance of R^(E) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, at least one instance of R^(E) isunsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, orhexyl). In certain embodiments, at least one instance of R^(E) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least one instance of R^(E) is methyl. In certain embodiments, atleast two instances of R^(E) are independently optionally substitutedalkyl. In certain embodiments, at least two instances of R^(E) areindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,at least two instances of R^(E) are independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, at least two instances of R^(E) are independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least two instances of R^(E) are methyl. In certain embodiments, z is2; and each instance of R^(E) is independently optionally substitutedalkyl. In certain embodiments, z is 2; and each instance of R^(E) isindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,z is 2; and each instance of R^(E) is independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, z is 2; and each instance of R^(E) is independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,z is 2; and each instance of R^(E) is methyl. In certain embodiments, zis 2; both instances of R^(E) are attached to the same carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; both instances of R^(E) are attached to thesame carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; both instances of R^(E) are attached to the samecarbon; and each instance of R^(E) is methyl. In certain embodiments, zis 2; each instance of R^(E) is attached to a different carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; each instance of R^(E) is attached to adifferent carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; each instance ofR^(E) is attached to a different carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; each instanceof R^(E) is attached to a different carbon; and each instance of R^(E)is independently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; each instance of R^(E) is attached to a differentcarbon; and each instance of R^(E) is methyl. In certain embodiments,two R^(E) groups are joined to form an optionally substitutedcarbocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl). In certain embodiments, two R^(E) groups are joined to forman optionally substituted cyclopropyl ring. In certain embodiments, twoR^(E) groups are joined to form an unsubstituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an unsubstitutedcyclopropyl ring. In certain embodiments, L₁ is a bond and R^(W) ishydrogen. In certain embodiments, L₁ is a bond, R^(W) is hydrogen, andR⁵ is hydrogen. In certain embodiments, L₁ is a bond and R^(W) ishalogen, e.g., fluoro, chloro, bromo, or iodo. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl, e.g.,unsubstituted C₁₋₄alkyl, e.g., methyl, ethyl, propyl, butyl, isopropyl,isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-214), provided arecompounds of Formulae XII-q1 to XII-q5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, z and R^(E) are defined herein. In certainembodiments, R³ is hydrogen. In certain embodiments, R³ is unsubstitutedC₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). In certainembodiments, R³ is methyl. In certain embodiments, R⁴ is hydrogen. Incertain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g., methyl,ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl. Incertain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, z is 2. In certain embodiments, at least oneinstance of R^(E) is optionally substituted alkyl. In certainembodiments, at least one instance of R^(E) is optionally substitutedC₁₋₆ alkyl. In certain embodiments, at least one instance of R^(E) isunsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, orhexyl). In certain embodiments, at least one instance of R^(E) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least one instance of R^(E) is methyl. In certain embodiments, atleast two instances of R^(E) are independently optionally substitutedalkyl. In certain embodiments, at least two instances of R^(E) areindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,at least two instances of R^(E) are independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, at least two instances of R^(E) are independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,at least two instances of R^(E) are methyl. In certain embodiments, z is2; and each instance of R^(E) is independently optionally substitutedalkyl. In certain embodiments, z is 2; and each instance of R^(E) isindependently optionally substituted C₁₋₆ alkyl. In certain embodiments,z is 2; and each instance of R^(E) is independently unsubstituted C₁₋₆alkyl (e.g., methyl, ethyl, propyl, butyl, pentyl, or hexyl). In certainembodiments, z is 2; and each instance of R^(E) is independentlyunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,z is 2; and each instance of R^(E) is methyl. In certain embodiments, zis 2; both instances of R^(E) are attached to the same carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; both instances of R^(E) are attached to thesame carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; both instancesof R^(E) are attached to the same carbon; and each instance of R^(E) isindependently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; both instances of R^(E) are attached to the samecarbon; and each instance of R^(E) is methyl. In certain embodiments, zis 2; each instance of R^(E) is attached to a different carbon; and eachinstance of R^(E) is independently optionally substituted alkyl. Incertain embodiments, z is 2; each instance of R^(E) is attached to adifferent carbon; and each instance of R^(E) is independently optionallysubstituted C₁₋₆ alkyl. In certain embodiments, z is 2; each instance ofR^(E) is attached to a different carbon; and each instance of R^(E) isindependently unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl, propyl,butyl, pentyl, or hexyl). In certain embodiments, z is 2; each instanceof R^(E) is attached to a different carbon; and each instance of R^(E)is independently unsubstituted branched C₃₋₆ alkyl (e.g., isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certainembodiments, z is 2; each instance of R^(E) is attached to a differentcarbon; and each instance of R^(E) is methyl. In certain embodiments,two R^(E) groups are joined to form an optionally substitutedcarbocyclic ring (e.g., cyclopropyl, cyclobutyl, cyclopentyl, orcyclohexyl). In certain embodiments, two R^(E) groups are joined to forman optionally substituted cyclopropyl ring. In certain embodiments, twoR^(E) groups are joined to form an unsubstituted carbocyclic ring (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl). In certainembodiments, two R^(E) groups are joined to form an unsubstitutedcyclopropyl ring. In certain embodiments, L₁ is a bond and R^(W) ishydrogen. In certain embodiments, L₁ is a bond, R^(W) is hydrogen, andR⁵ is hydrogen. In certain embodiments, L₁ is a bond and R^(W) ishalogen, e.g., fluoro, chloro, bromo, or iodo. In certain embodiments,L₁ is a bond and R^(W) is optionally substituted C₁₋₆alkyl, e.g.,unsubstituted C₁₋₄alkyl, e.g., methyl, ethyl, propyl, butyl, isopropyl,isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-222), provided arecompounds of Formulae XII-r1 to XII-r5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, z, R^(E), R^(G), and R^(H) are defined herein. Incertain embodiments, R³ is hydrogen. In certain embodiments, R³ isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, R³ is methyl. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g.,methyl, ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl.In certain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, z is O. In certain embodiments, at least oneinstance of R^(G) is hydrogen. In certain embodiments, both instances ofR^(G) are hydrogen. In certain embodiments, R^(H) is hydrogen. Incertain embodiments, R^(H) is optionally substituted alkyl. In certainembodiments, R^(H) is optionally substituted C₁₋₆ alkyl. In certainembodiments, R^(H) is unsubstituted C₁₋₆ alkyl (e.g., methyl, ethyl,propyl, butyl, pentyl, or hexyl). In certain embodiments, R^(H) isunsubstituted branched C₃₋₆ alkyl (e.g., isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or 3-pentyl). In certain embodiments,R^(H) is methyl. In certain embodiments, R^(H) is ethyl. In certainembodiments, R^(H) is isopropyl. In certain embodiments, R^(H) istert-butyl. In certain embodiments, R^(H) is optionally substitutedcarbocyclyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl).In certain embodiments, R^(H) is optionally substituted aryl. In certainembodiments, R^(H) is optionally substituted heterocyclyl. In certainembodiments, R^(H) is optionally substituted heteroaryl. In certainembodiments, two R^(E) groups are joined to form an optionallysubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an optionally substituted cyclopropyl ring. Incertain embodiments, two R^(E) groups are joined to form anunsubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an unsubstituted cyclopropyl ring. In certainembodiments, L₁ is a bond and R^(W) is hydrogen. In certain embodiments,L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. In certainembodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro, chloro,bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W) isoptionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl, e.g.,methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, when Ring A is of Formula (r-231), provided arecompounds of Formulae XII-s1 to XII-s5:

or pharmaceutically acceptable salt thereof; wherein R³, R⁴, R⁵, R^(x),V₂₁, V₂₂, V₂₃, V₂₄, z, R^(E), R^(G), and R^(Ha) are defined herein. Incertain embodiments, R³ is hydrogen. In certain embodiments, R³ isunsubstituted C₁₋₄ alkyl (e.g., methyl, ethyl, propyl, or butyl). Incertain embodiments, R³ is methyl. In certain embodiments, R⁴ ishydrogen. In certain embodiments, R⁴ is unsubstituted C₁₋₆ alkyl (e.g.,methyl, ethyl, propyl, or butyl). In certain embodiments, R⁴ is methyl.In certain embodiments, R^(x) is unsubstituted C₁₋₄ alkyl (e.g., methyl,ethyl, propyl or butyl). In certain embodiments, R^(x) is methyl. Incertain embodiments, R⁵ is hydrogen. In certain embodiments, at leastone of V₂₁, V₂₂, V₂₃, and V₂₄ is —CH₂—. In certain embodiments, at leasttwo of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast three of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments,all four of V₂₁, V₂₂, V₂₃, and V₂₄ are —CH₂—. In certain embodiments, atleast one of V₂₃ and V₂₄ is O or —CH(OR^(A))—. In certain embodiments,V₂₃ is O. In certain embodiments, V₂₃ is —CH(OR^(A))—. In certainembodiments, V₂₄ is O. In certain embodiments, V₂₄ is —CH(OR^(A))—. Incertain embodiments, z is O. In certain embodiments, at least oneinstance of R^(G) is hydrogen. In certain embodiments, both instances ofR^(G) are hydrogen. In certain embodiments, R^(Ha) is halogen (e.g.,fluorine, chlorine, or bromine). In certain embodiments, R^(Ha) isfluorine. In certain embodiments, two R^(E) groups are joined to form anoptionally substituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an optionally substituted cyclopropyl ring. Incertain embodiments, two R^(E) groups are joined to form anunsubstituted carbocyclic ring (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl). In certain embodiments, two R^(E) groupsare joined to form an unsubstituted cyclopropyl ring. In certainembodiments, L₁ is a bond and R^(W) is hydrogen. In certain embodiments,L₁ is a bond, R^(W) is hydrogen, and R⁵ is hydrogen. In certainembodiments, L₁ is a bond and R^(W) is halogen, e.g., fluoro, chloro,bromo, or iodo. In certain embodiments, L₁ is a bond and R^(W) isoptionally substituted C₁₋₆alkyl, e.g., unsubstituted C₁₋₄alkyl, e.g.,methyl, ethyl, propyl, butyl, isopropyl, isobutyl, or isoamyl.

In certain embodiments, a provided compound is a compound listed inTable 1A, Table 1B, or a pharmaceutically acceptable salt thereof.

TABLE 1A Exemplary Compounds LC-MS Cmpd No Structure m/z (M + H) 1

299.30 2

325.30 3

269.30 4

287.20 5

347.00 6

339.20 7

301.20 8

271.20 9

259.10 10

237.10 11

317.30 12

195.10 13

293.10 14

393.10 15

293.10 16

325.10 17

287.30 18

325.10 19

225.20 20

— 21

197.30 22

237.10 23

277.05 24

271.40 25

221.30 26

209.50 27

211.40 28

211.70 29

197.30 30

169.80 31

223.30 32

370.00 33

237.05 34

322.15 35

251.00 36

265.05 37

251.10 38

265.10 39

308.20 40

336.15 41

266.15 42

252.10 43

322.15 44

322.15

TABLE 1B Exemplary Compounds Cmpd LC-MS m/z No Structure (M + H) 45

291.15 46

319.15 47

319.15 48

305.15 49

319.15 50

293.1 51

307.10 52

279.10 53

295.1 54

307.1 55

309.1 56

309.15 57

339.10 58

325.15 59

309.25 60

427.25 61

395.3 62

— 63

349.25 64

— 65

— 66

— 67

343.15 68

— 69

— 70

337.2 71

— 72

— 73

— 74

— 75

— 76

— 77

— 78

— 79

— 80

— 81

— 82

— 83

— 84

— 85

— 86

— 87

— 88

— 89

— 90

— 91

— 92

— 93

— 94

— 95

— 96

— 97

— 98

— 99

— 100

— 101

— 102

— 103

— 104

— 105

— 106

— 107

448.4 108

420.2 109

420.2 110

370.1 111

484.2 112

339.15 113

369.25 114

448.3 115

356.15 116

461.4 117

336.2 118

367.3 119

367.3 120

484.35 121

351.25 122

395.25 123

428.3 124

376.3 125

428.3 126

446.3 127

410.25 128

410.3 129

376.3 130

355.25 131

367.3 132

339.2 133

335.2 134

335.2 135

311.25 136

387.2 137

321.1 138

339.1 139

349.25 140

349.25 141

307.15 142

307.15 143

307.25 144

277.15 145

391.15 146

440.15 147

320.1 148

279.25 149

265.1 150

365.25 151

391.3 152

334.15 153

323.15 154

363.15 155

376.25 156

376.25 157

334.15 158

367.2 159

407.35 160

323.2 161

349 162

363.05 163

390.3 164

376.25 165

390.3 166

295.1 167

295.1 168

393.3 169

307.15 170

349.15 171

363.2 172

349.2 173

365.2 174

367.35 175

335.1 176

320.1 177

309.1 178

321.05 179

281.1 180

293.15 181

293.15 182

321.3 183

— 184

— 185

295.05 186

417.10 187

387.10 188

367.2 189

367.2 190

338.1 191

424.2 192

448.2 193

448.4 194

265.1 195

420.4 196

420.5 197

446.2 198

309.2 199

307.15 200

335.1 201

335.2 202

337.1 203

364.2 204

364.2 205

349.2 206

349.15 207

321.15 208

321.1 209

378.2 210

378.2 211

414.1 212

414.1 213

436.2 214

436.2 215

364.2 216

287.05 217

267.05 218

472.4 219

472.4 220

442.2 221

442.2 222

470.2 223

470.4 224

442.3 225

442.3 226

364.2 227

267.05 228

486.4 229

486.4 230

472.4 231

470.2 232

470.2 233

408.2 234

408.2 235

422.4 236

422.4 237

436.4 238

436.4 239

378.3 240

378.3 241

422.2 242

422.2 243

436.2 244

436.2 245

349.1 246

486.3 247

486.3 248

422.3 249

422.3 250

392.2 251

392.3 252

408.2 253

335.2 254

422.3 255

— 256

337.5 257

349.2 258

428.1 259

428.1 260

335.1 261

422.3 262

378.2 263

392.2 264

380.2 265

394.2 266

378.15 267

505.2 268

380.3 269

380.3 270

394.3 271

394.5 272

367.15 273

393.21 274

335.1 275

305.4

In certain embodiments, a provided compound inhibits an RMT (e.g.,PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8). In certain embodiments, aprovided compound inhibits wild-type PRMT1, PRMT3, CARM1, PRMT6, and/orPRMT8. In certain embodiments, a provided compound inhibits a mutantRMT. In certain embodiments, a provided compound inhibits PRMT1, PRMT3,CARM1, PRMT6, and/or PRMT8, e.g., as measured in an assay describedherein. In certain embodiments, the RMT is from a human. In certainembodiments, a provided compound inhibits an RMT (e.g., PRMT1, PRMT3,CARM1, PRMT6, and/or PRMT8) at an IC₅₀ less than or equal to 10 μM. Incertain embodiments, a provided compound inhibits an RMT (e.g., PRMT1,PRMT3, CARM1, PRMT6, and/or PRMT8) at an IC₅₀ less than or equal to 1μM. In certain embodiments, a provided compound inhibits an RMT (e.g.,PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8) at an IC₅₀ less than or equalto 0.1 μM. In certain embodiments, a provided compound inhibits an RMT(e.g., PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8) at an IC50 less than orequal to 0.01 μM. In certain embodiments, a provided compound inhibitsan RMT (e.g., PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8) in a cell at anEC₃₀ less than or equal to 10 μM. In certain embodiments, a providedcompound inhibits an RMT (e.g., PRMT1, PRMT3, CARM1, PRMT6, and/orPRMT8) in a cell at an EC₃₀ less than or equal to 12 μM. In certainembodiments, a provided compound inhibits an RMT (e.g., PRMT1, PRMT3,CARM1, PRMT6, and/or PRMT8) in a cell at an EC₃₀ less than or equal to 3μM. In certain embodiments, a provided compound inhibits PRMT1 in a cellat an EC₃₀ less than or equal to 12 μM. In certain embodiments, aprovided compound inhibits PRMT1 in a cell at an EC₃₀ less than or equalto 3 μM. In certain embodiments, a provided compound inhibits an RMT(e.g., PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8) in a cell at an EC₃₀less than or equal to 1 μM. In certain embodiments, a provided compoundinhibits an RMT (e.g., PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8) in acell at an EC₃₀ less than or equal to 0.1 μM. In certain embodiments, aprovided compound inhibits cell proliferation at an EC₅₀ less than orequal to 10 μM. In certain embodiments, a provided compound inhibitscell proliferation at an EC₅₀ less than or equal to 1 μM. In certainembodiments, a provided compound inhibits cell proliferation at an EC₅₀less than or equal to 0.1 μM.

It will be understood by one of ordinary skill in the art that the RMTcan be wild-type, or any mutant or variant.

The present disclosure provides pharmaceutical compositions comprising acompound described herein, e.g., a compound of Formula (I), or apharmaceutically acceptable salt thereof, as described herein, andoptionally a pharmaceutically acceptable excipient. It will beunderstood by one of ordinary skill in the art that the compoundsdescribed herein, or salts thereof, may be present in various forms,such as amorphous, hydrates, solvates, or polymorphs. In certainembodiments, a provided composition comprises two or more compoundsdescribed herein. In certain embodiments, a compound described herein,or a pharmaceutically acceptable salt thereof, is provided in aneffective amount in the pharmaceutical composition. In certainembodiments, the effective amount is a therapeutically effective amount.In certain embodiments, the effective amount is an amount effective forinhibiting an RMT (e.g., PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8). Incertain embodiments, the effective amount is an amount effective fortreating an RMT-mediated disorder (e.g., a PRMT1-, PRMT3-, CARM1-,PRMT6-, and/or PRMT8-mediated disorder). In certain embodiments, theeffective amount is a prophylactically effective amount. In certainembodiments, the effective amount is an amount effective to prevent anRMT-mediated disorder.

Pharmaceutically acceptable excipients include any and all solvents,diluents, or other liquid vehicles, dispersions, suspension aids,surface active agents, isotonic agents, thickening or emulsifyingagents, preservatives, solid binders, lubricants, and the like, assuited to the particular dosage form desired. General considerations informulation and/or manufacture of pharmaceutical compositions agents canbe found, for example, in Remington's Pharmaceutical Sciences, SixteenthEdition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980), andRemington: The Science and Practice of Pharmacy, 21st Edition(Lippincott Williams & Wilkins, 2005).

Pharmaceutical compositions described herein can be prepared by anymethod known in the art of pharmacology. In general, such preparatorymethods include the steps of bringing a compound described herein (the“active ingredient”) into association with a carrier and/or one or moreother accessory ingredients, and then, if necessary and/or desirable,shaping and/or packaging the product into a desired single- ormulti-dose unit.

Pharmaceutical compositions can be prepared, packaged, and/or sold inbulk, as a single unit dose, and/or as a plurality of single unit doses.As used herein, a “unit dose” is discrete amount of the pharmaceuticalcomposition comprising a predetermined amount of the active ingredient.The amount of the active ingredient is generally equal to the dosage ofthe active ingredient which would be administered to a subject and/or aconvenient fraction of such a dosage such as, for example, one-half orone-third of such a dosage.

Relative amounts of the active ingredient, the pharmaceuticallyacceptable excipient, and/or any additional ingredients in apharmaceutical composition of the present disclosure will vary,depending upon the identity, size, and/or condition of the subjecttreated and further depending upon the route by which the composition isto be administered. By way of example, the composition may comprisebetween 0.1% and 100% (w/w) active ingredient.

In some embodiments, a pharmaceutical composition described herein issterilized.

Pharmaceutically acceptable excipients used in the manufacture ofprovided pharmaceutical compositions include inert diluents, dispersingand/or granulating agents, surface active agents and/or emulsifiers,disintegrating agents, binding agents, preservatives, buffering agents,lubricating agents, and/or oils. Excipients such as cocoa butter andsuppository waxes, coloring agents, coating agents, sweetening,flavoring, and perfuming agents may also be present in the composition.

Exemplary diluents include calcium carbonate, sodium carbonate, calciumphosphate, dicalcium phosphate, calcium sulfate, calcium hydrogenphosphate, sodium phosphate lactose, sucrose, cellulose,microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodiumchloride, dry starch, cornstarch, powdered sugar, and mixtures thereof.

Exemplary granulating and/or dispersing agents include potato starch,corn starch, tapioca starch, sodium starch glycolate, clays, alginicacid, guar gum, citrus pulp, agar, bentonite, cellulose and woodproducts, natural sponge, cation-exchange resins, calcium carbonate,silicates, sodium carbonate, cross-linked poly(vinyl-pyrrolidone)(crospovidone), sodium carboxymethyl starch (sodium starch glycolate),carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose(croscarmellose), methylcellulose, pregelatinized starch (starch 1500),microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate,quaternary ammonium compounds, and mixtures thereof.

Exemplary surface active agents and/or emulsifiers include naturalemulsifiers (e.g., acacia, agar, alginic acid, sodium alginate,tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk,casein, wool fat, cholesterol, wax, and lecithin), colloidal clays(e.g., bentonite (aluminum silicate) and Veegum (magnesium aluminumsilicate)), long chain amino acid derivatives, high molecular weightalcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetinmonostearate, ethylene glycol distearate, glyceryl monostearate, andpropylene glycol monostearate, polyvinyl alcohol), carbomers (e.g.,carboxy polymethylene, polyacrylic acid, acrylic acid polymer, andcarboxyvinyl polymer), carrageenan, cellulosic derivatives (e.g.,carboxymethylcellulose sodium, powdered cellulose, hydroxymethylcellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose,methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylenesorbitan monolaurate (Tween 20), polyoxyethylene sorbitan (Tween 60),polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monopalmitate(Span 40), sorbitan monostearate (Span 60], sorbitan tristearate (Span65), glyceryl monooleate, sorbitan monooleate (Span 80)),polyoxyethylene esters (e.g., polyoxyethylene monostearate (Myrj 45),polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil,polyoxymethylene stearate, and Solutol), sucrose fatty acid esters,polyethylene glycol fatty acid esters (e.g., Cremophor™),polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether (Brij 30)),poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamineoleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyllaurate, sodium lauryl sulfate, Pluronic F68, Poloxamer 188, cetrimoniumbromide, cetylpyridinium chloride, benzalkonium chloride, docusatesodium, and/or mixtures thereof.

Exemplary binding agents include starch (e.g., cornstarch and starchpaste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin,molasses, lactose, lactitol, mannitol, etc.), natural and synthetic gums(e.g., acacia, sodium alginate, extract of Irish moss, panwar gum,ghatti gum, mucilage of isapol husks, carboxymethylcellulose,methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose,cellulose acetate, poly(vinyl-pyrrolidone), magnesium aluminum silicate(Veegum), and larch arabogalactan), alginates, polyethylene oxide,polyethylene glycol, inorganic calcium salts, silicic acid,polymethacrylates, waxes, water, alcohol, and/or mixtures thereof.

Exemplary preservatives include antioxidants, chelating agents,antimicrobial preservatives, antifungal preservatives, alcoholpreservatives, acidic preservatives, and other preservatives.

Exemplary antioxidants include alpha tocopherol, ascorbic acid, acorbylpalmitate, butylated hydroxyanisole, butylated hydroxytoluene,monothioglycerol, potassium metabisulfite, propionic acid, propylgallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, andsodium sulfite.

Exemplary chelating agents include ethylenediaminetetraacetic acid(EDTA) and salts and hydrates thereof (e.g., sodium edetate, disodiumedetate, trisodium edetate, calcium disodium edetate, dipotassiumedetate, and the like), citric acid and salts and hydrates thereof(e.g., citric acid monohydrate), fumaric acid and salts and hydratesthereof, malic acid and salts and hydrates thereof, phosphoric acid andsalts and hydrates thereof, and tartaric acid and salts and hydratesthereof. Exemplary antimicrobial preservatives include benzalkoniumchloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide,cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol,chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea,phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate,propylene glycol, and thimerosal.

Exemplary antifungal preservatives include butyl paraben, methylparaben, ethyl paraben, propyl paraben, benzoic acid, hydroxybenzoicacid, potassium benzoate, potassium sorbate, sodium benzoate, sodiumpropionate, and sorbic acid.

Exemplary alcohol preservatives include ethanol, polyethylene glycol,phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate,and phenylethyl alcohol. Exemplary acidic preservatives include vitaminA, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid,dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.

Other preservatives include tocopherol, tocopherol acetate, deteroximemesylate, cetrimide, butylated hydroxyanisol (BHA), butylatedhydroxytoluened (BHT), ethylenediamine, sodium lauryl sulfate (SLS),sodium lauryl ether sulfate (SLES), sodium bisulfite, sodiummetabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus,Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, andEuxyl. In certain embodiments, the preservative is an anti-oxidant. Inother embodiments, the preservative is a chelating agent.

Exemplary buffering agents include citrate buffer solutions, acetatebuffer solutions, phosphate buffer solutions, ammonium chloride, calciumcarbonate, calcium chloride, calcium citrate, calcium glubionate,calcium gluceptate, calcium gluconate, D-gluconic acid, calciumglycerophosphate, calcium lactate, propanoic acid, calcium levulinate,pentanoic acid, dibasic calcium phosphate, phosphoric acid, tribasiccalcium phosphate, calcium hydroxide phosphate, potassium acetate,potassium chloride, potassium gluconate, potassium mixtures, dibasicpotassium phosphate, monobasic potassium phosphate, potassium phosphatemixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodiumcitrate, sodium lactate, dibasic sodium phosphate, monobasic sodiumphosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide,aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline,Ringer's solution, ethyl alcohol, and mixtures thereof.

Exemplary lubricating agents include magnesium stearate, calciumstearate, stearic acid, silica, talc, malt, glyceryl behanate,hydrogenated vegetable oils, polyethylene glycol, sodium benzoate,sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate,sodium lauryl sulfate, and mixtures thereof.

Exemplary natural oils include almond, apricot kernel, avocado, babassu,bergamot, black current seed, borage, cade, camomile, canola, caraway,camauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee,corn, cotton seed, emu, eucalyptus, evening primrose, fish, flaxseed,geraniol, gourd, grape seed, hazel nut, hyssop, isopropyl myristate,jojoba, kukui nut, lavandin, lavender, lemon, litsea cubeba, macademianut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange,orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed,pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood,sasquana, savoury, sea buckthom, sesame, shea butter, silicone, soybean,sunflower, tea tree, thistle, tsubaki, vetiver, walnut, and wheat germoils. Exemplary synthetic oils include, but are not limited to, butylstearate, caprylic triglyceride, capric triglyceride, cyclomethicone,diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil,octyldodecanol, oleyl alcohol, silicone oil, and mixtures thereof.

Liquid dosage forms for oral and parenteral administration includepharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active ingredients,the liquid dosage forms may comprise inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed,groundnut, corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can include adjuvants such as wetting agents, emulsifyingand suspending agents, sweetening, flavoring, and perfuming agents. Incertain embodiments for parenteral administration, the compoundsdescribed herein are mixed with solubilizing agents such as Cremophor™,alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins,polymers, and mixtures thereof.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions can be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation can be a sterile injectable solution,suspension or emulsion in a nontoxic parenterally acceptable diluent orsolvent, for example, as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that can be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This can be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Compositions for rectal or vaginal administration are typicallysuppositories which can be prepared by mixing the compounds describedherein with suitable non-irritating excipients or carriers such as cocoabutter, polyethylene glycol or a suppository wax which are solid atambient temperature but liquid at body temperature and therefore melt inthe rectum or vaginal cavity and release the active ingredient.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activeingredient is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may comprise buffering agents.

Solid compositions of a similar type can be employed as fillers in softand hard-filled gelatin capsules using such excipients as lactose ormilk sugar as well as high molecular weight polyethylene glycols and thelike. The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes. Solid compositions of asimilar type can be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polyethylene glycols and the like.

The active ingredient can be in micro-encapsulated form with one or moreexcipients as noted above. The solid dosage forms of tablets, dragees,capsules, pills, and granules can be prepared with coatings and shellssuch as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active ingredient can be admixed with at least oneinert diluent such as sucrose, lactose, or starch. Such dosage forms maycomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets, and pills, the dosage forms may comprise bufferingagents. They may optionally comprise opacifying agents and can be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions which can beused include polymeric substances and waxes.

Dosage forms for topical and/or transdermal administration of a providedcompound may include ointments, pastes, creams, lotions, gels, powders,solutions, sprays, inhalants and/or patches. Generally, the activeingredient is admixed under sterile conditions with a pharmaceuticallyacceptable carrier and/or any desired preservatives and/or buffers ascan be required. Additionally, the present disclosure encompasses theuse of transdermal patches, which often have the added advantage ofproviding controlled delivery of an active ingredient to the body. Suchdosage forms can be prepared, for example, by dissolving and/ordispensing the active ingredient in the proper medium. Alternatively oradditionally, the rate can be controlled by either providing a ratecontrolling membrane and/or by dispersing the active ingredient in apolymer matrix and/or gel.

Formulations suitable for topical administration include, but are notlimited to, liquid and/or semi liquid preparations such as liniments,lotions, oil in water and/or water in oil emulsions such as creams,ointments and/or pastes, and/or solutions and/or suspensions.Topically-administrable formulations may, for example, comprise fromabout 1% to about 10% (w/w) active ingredient, although theconcentration of the active ingredient can be as high as the solubilitylimit of the active ingredient in the solvent. Formulations for topicaladministration may further comprise one or more of the additionalingredients described herein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation suitable for pulmonary administration via thebuccal cavity. Such a formulation may comprise dry particles whichcomprise the active ingredient and which have a diameter in the rangefrom about 0.5 to about 7 nanometers or from about 1 to about 6nanometers. Such compositions are conveniently in the form of drypowders for administration using a device comprising a dry powderreservoir to which a stream of propellant can be directed to dispersethe powder and/or using a self propelling solvent/powder dispensingcontainer such as a device comprising the active ingredient dissolvedand/or suspended in a low-boiling propellant in a sealed container. Suchpowders comprise particles wherein at least 98% of the particles byweight have a diameter greater than 0.5 nanometers and at least 95% ofthe particles by number have a diameter less than 7 nanometers.Alternatively, at least 95% of the particles by weight have a diametergreater than 1 nanometer and at least 90% of the particles by numberhave a diameter less than 6 nanometers. Dry powder compositions mayinclude a solid fine powder diluent such as sugar and are convenientlyprovided in a unit dose form.

Low boiling propellants generally include liquid propellants having aboiling point of below 65° F. at atmospheric pressure. Generally thepropellant may constitute 50 to 99.9% (w/w) of the composition, and theactive ingredient may constitute 0.1 to 20% (w/w) of the composition.The propellant may further comprise additional ingredients such as aliquid non-ionic and/or solid anionic surfactant and/or a solid diluent(which may have a particle size of the same order as particlescomprising the active ingredient).

Pharmaceutical compositions formulated for pulmonary delivery mayprovide the active ingredient in the form of droplets of a solutionand/or suspension. Such formulations can be prepared, packaged, and/orsold as aqueous and/or dilute alcoholic solutions and/or suspensions,optionally sterile, comprising the active ingredient, and mayconveniently be administered using any nebulization and/or atomizationdevice. Such formulations may further comprise one or more additionalingredients including, but not limited to, a flavoring agent such assaccharin sodium, a volatile oil, a buffering agent, a surface activeagent, and/or a preservative such as methylhydroxybenzoate. The dropletsprovided by this route of administration may have an average diameter inthe range from about 0.1 to about 200 nanometers.

Formulations described herein as being useful for pulmonary delivery areuseful for intranasal delivery of a pharmaceutical composition. Anotherformulation suitable for intranasal administration is a coarse powdercomprising the active ingredient and having an average particle fromabout 0.2 to 500 micrometers. Such a formulation is administered byrapid inhalation through the nasal passage from a container of thepowder held close to the nares.

Formulations for nasal administration may, for example, comprise fromabout as little as 0.1% (w/w) and as much as 100% (w/w) of the activeingredient, and may comprise one or more of the additional ingredientsdescribed herein. A provided pharmaceutical composition can be prepared,packaged, and/or sold in a formulation for buccal administration. Suchformulations may, for example, be in the form of tablets and/or lozengesmade using conventional methods, and may contain, for example, 0.1 to20% (w/w) active ingredient, the balance comprising an orallydissolvable and/or degradable composition and, optionally, one or moreof the additional ingredients described herein. Alternately,formulations for buccal administration may comprise a powder and/or anaerosolized and/or atomized solution and/or suspension comprising theactive ingredient. Such powdered, aerosolized, and/or aerosolizedformulations, when dispersed, may have an average particle and/ordroplet size in the range from about 0.1 to about 200 nanometers, andmay further comprise one or more of the additional ingredients describedherein.

A provided pharmaceutical composition can be prepared, packaged, and/orsold in a formulation for ophthalmic administration. Such formulationsmay, for example, be in the form of eye drops including, for example, a0.1/1.0% (w/w) solution and/or suspension of the active ingredient in anaqueous or oily liquid carrier. Such drops may further comprisebuffering agents, salts, and/or one or more other of the additionalingredients described herein. Other opthalmically-administrableformulations which are useful include those which comprise the activeingredient in microcrystalline form and/or in a liposomal preparation.Ear drops and/or eye drops are contemplated as being within the scope ofthis disclosure.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to animals of all sorts. Modification of pharmaceuticalcompositions suitable for administration to humans in order to renderthe compositions suitable for administration to various animals is wellunderstood, and the ordinarily skilled veterinary pharmacologist candesign and/or perform such modification with ordinary experimentation.

Compounds provided herein are typically formulated in dosage unit formfor ease of administration and uniformity of dosage. It will beunderstood, however, that the total daily usage of provided compositionswill be decided by the attending physician within the scope of soundmedical judgment. The specific therapeutically effective dose level forany particular subject or organism will depend upon a variety of factorsincluding the disease, disorder, or condition being treated and theseverity of the disorder; the activity of the specific active ingredientemployed; the specific composition employed; the age, body weight,general health, sex and diet of the subject; the time of administration,route of administration, and rate of excretion of the specific activeingredient employed; the duration of the treatment; drugs used incombination or coincidental with the specific active ingredientemployed; and like factors well known in the medical arts.

The compounds and compositions provided herein can be administered byany route, including enteral (e.g., oral), parenteral, intravenous,intramuscular, intra-arterial, intramedullary, intrathecal,subcutaneous, intraventricular, transdermal, interdermal, rectal,intravaginal, intraperitoneal, topical (as by powders, ointments,creams, and/or drops), mucosal, nasal, bucal, sublingual; byintratracheal instillation, bronchial instillation, and/or inhalation;and/or as an oral spray, nasal spray, and/or aerosol. Specificallycontemplated routes are oral administration, intravenous administration(e.g., systemic intravenous injection), regional administration viablood and/or lymph supply, and/or direct administration to an affectedsite. In general the most appropriate route of administration willdepend upon a variety of factors including the nature of the agent(e.g., its stability in the environment of the gastrointestinal tract),and/or the condition of the subject (e.g., whether the subject is ableto tolerate oral administration).

The exact amount of a compound required to achieve an effective amountwill vary from subject to subject, depending, for example, on species,age, and general condition of a subject, severity of the side effects ordisorder, identity of the particular compound(s), mode ofadministration, and the like. The desired dosage can be delivered threetimes a day, two times a day, once a day, every other day, every thirdday, every week, every two weeks, every three weeks, or every fourweeks. In certain embodiments, the desired dosage can be delivered usingmultiple administrations (e.g., two, three, four, five, six, seven,eight, nine, ten, eleven, twelve, thirteen, fourteen, or moreadministrations).

In certain embodiments, an effective amount of a compound foradministration one or more times a day to a 70 kg adult human maycomprise about 0.0001 mg to about 3000 mg, about 0.0001 mg to about 2000mg, about 0.0001 mg to about 1000 mg, about 0.001 mg to about 1000 mg,about 0.01 mg to about 1000 mg, about 0.1 mg to about 1000 mg, about 1mg to about 1000 mg, about 1 mg to about 100 mg, about 10 mg to about1000 mg, or about 100 mg to about 1000 mg, of a compound per unit dosageform.

In certain embodiments, a compound described herein may be administeredat dosage levels sufficient to deliver from about 0.001 mg/kg to about1000 mg/kg, from about 0.01 mg/kg to about mg/kg, from about 0.1 mg/kgto about 40 mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about0.01 mg/kg to about 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, orfrom about 1 mg/kg to about 25 mg/kg, of subject body weight per day,one or more times a day, to obtain the desired therapeutic effect.

In some embodiments, a compound described herein is administered one ormore times per day, for multiple days. In some embodiments, the dosingregimen is continued for days, weeks, months, or years.

It will be appreciated that dose ranges as described herein provideguidance for the administration of provided pharmaceutical compositionsto an adult. The amount to be administered to, for example, a child oran adolescent can be determined by a medical practitioner or personskilled in the art and can be lower or the same as that administered toan adult.

It will be also appreciated that a compound or composition, as describedherein, can be administered in combination with one or more additionaltherapeutically active agents. In certain embodiments, a compound orcomposition provided herein is administered in combination with one ormore additional therapeutically active agents that improve itsbioavailability, reduce and/or modify its metabolism, inhibit itsexcretion, and/or modify its distribution within the body. It will alsobe appreciated that the therapy employed may achieve a desired effectfor the same disorder, and/or it may achieve different effects.

The compound or composition can be administered concurrently with, priorto, or subsequent to, one or more additional therapeutically activeagents. In certain embodiments, the additional therapeutically activeagent is a compound of Formula (I). In certain embodiments, theadditional therapeutically active agent is not a compound of Formula(I). In general, each agent will be administered at a dose and/or on atime schedule determined for that agent. In will further be appreciatedthat the additional therapeutically active agent utilized in thiscombination can be administered together in a single composition oradministered separately in different compositions. The particularcombination to employ in a regimen will take into account compatibilityof a provided compound with the additional therapeutically active agentand/or the desired therapeutic effect to be achieved. In general, it isexpected that additional therapeutically active agents utilized incombination be utilized at levels that do not exceed the levels at whichthey are utilized individually. In some embodiments, the levels utilizedin combination will be lower than those utilized individually.

Exemplary additional therapeutically active agents include, but are notlimited to, small organic molecules such as drug compounds (e.g.,compounds approved by the U.S. Food and Drug Administration as providedin the Code of Federal Regulations (CFR)), peptides, proteins,carbohydrates, monosaccharides, oligosaccharides, polysaccharides,nucleoproteins, mucoproteins, lipoproteins, synthetic polypeptides orproteins, small molecules linked to proteins, glycoproteins, steroids,nucleic acids, DNAs, RNAs, nucleotides, nucleosides, oligonucleotides,antisense oligonucleotides, lipids, hormones, vitamins, and cells. Incertain embodiments, an additional therapeutically active agent isprednisolone, dexamethasone, doxorubicin, vincristine, mafosfamide,cisplatin, carboplatin, Ara-C, rituximab, azacitadine, panobinostat,vorinostat, everolimus, rapamycin, ATRA (all-trans retinoic acid),daunorubicin, decitabine, Vidaza, mitoxantrone, or IBET-151.

Also encompassed by the present disclosure are kits (e.g.,pharmaceutical packs). The kits provided may comprise a providedpharmaceutical composition or compound and a container (e.g., a vial,ampule, bottle, syringe, and/or dispenser package, or other suitablecontainer). In some embodiments, provided kits may optionally furtherinclude a second container comprising a pharmaceutical excipient fordilution or suspension of a provided pharmaceutical composition orcompound. In some embodiments, a provided pharmaceutical composition orcompound provided in the container and the second container are combinedto form one unit dosage form. In some embodiments, a provided kitsfurther includes instructions for use.

Compounds and compositions described herein are generally useful for theinhibition of RMT (e.g., PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8). Insome embodiments, methods of treating an RMT-mediated disorder in asubject are provided which comprise administering an effective amount ofa compound described herein (e.g., a compound of Formula (I)), or apharmaceutically acceptable salt thereof), to a subject in need oftreatment. In certain embodiments, the effective amount is atherapeutically effective amount. In certain embodiments, the effectiveamount is a prophylactically effective amount. In certain embodiments,the subject is suffering from a RMT-mediated disorder. In certainembodiments, the subject is susceptible to a RMT-mediated disorder.

As used herein, the term “RMT-mediated disorder” means any disease,disorder, or other pathological condition in which an RMT (e.g., PRMT1,PRMT3, CARM1, PRMT6, and/or PRMT8) is known to play a role. Accordingly,in some embodiments, the present disclosure relates to treating orlessening the severity of one or more diseases in which an RMT is knownto play a role.

In some embodiments, the present disclosure provides a method ofinhibiting an RMT comprising contacting the RMT with an effective amountof a compound described herein (e.g., a compound of Formula (I)), or apharmaceutically acceptable salt thereof. The RMT may be purified orcrude, and may be present in a cell, tissue, or subject. Thus, suchmethods encompass both inhibition of in vitro and in vivo RMT activity.In certain embodiments, the method is an in vitro method, e.g., such asan assay method. It will be understood by one of ordinary skill in theart that inhibition of an RMT does not necessarily require that all ofthe RMT be occupied by an inhibitor at once. Exemplary levels ofinhibition of an RMT (e.g., PRMT1, PRMT3, CARM1, PRMT6, and/or PRMT8)include at least 10% inhibition, about 10% to about 25% inhibition,about 25% to about 50% inhibition, about 50% to about 75% inhibition, atleast 50% inhibition, at least 75% inhibition, about 80% inhibition,about 90% inhibition, and greater than 90% inhibition.

In some embodiments, provided is a method of inhibiting RMT activity ina subject in need thereof (e.g., a subject diagnosed as having anRMT-mediated disorder) comprising administering to the subject aneffective amount of a compound described herein (e.g., a compound ofFormula (I)), or a pharmaceutically acceptable salt thereof, or apharmaceutical composition thereof.

In certain embodiments, provided is a method of modulating geneexpression in a cell which comprises contacting a cell with an effectiveamount of a compound of Formula (I), or a pharmaceutically acceptablesalt thereof. In certain embodiments, the cell is in culture in vitro.In certain embodiments, the cell is in an animal, e.g., a human. Incertain embodiments, the cell is in a subject in need of treatment.

In certain embodiments, provided is a method of modulating transcriptionin a cell which comprises contacting a cell with an effective amount ofa compound of Formula (I), or a pharmaceutically acceptable saltthereof. In certain embodiments, the cell is in culture in vitro. Incertain embodiments, the cell is in an animal, e.g., a human. In certainembodiments, the cell is in a subject in need of treatment.

In certain embodiments, a method is provided of selecting a therapy fora subject having a disease associated with an RMT-mediated disorder ormutation comprising the steps of determining the presence of anRMT-mediated disorder or gene mutation in an RMT gene (e.g., a PRMT1,PRMT3, CARM1, PRMT6, and/or PRMT8 gene) or and selecting, based on thepresence of an RMT-mediated disorder a gene mutation in the RMT gene atherapy that includes the administration of a provided compound. Incertain embodiments, the disease is cancer.

In certain embodiments, a method of treatment is provided for a subjectin need thereof comprising the steps of determining the presence of anRMT-mediated disorder or a gene mutation in the RMT gene and treatingthe subject in need thereof, based on the presence of a RMT-mediateddisorder or gene mutation in the RMT gene with a therapy that includesthe administration of a provided compound. In certain embodiments, thesubject is a cancer patient.

In some embodiments, a compound provided herein is useful in treating aproliferative disorder, such as cancer. For example, while not beingbound to any particular mechanism, protein arginine methylation by PRMTsis a modification that has been implicated in signal transduction, genetranscription, DNA repair and mRNA splicing, among others; andoverexpression of PRMTs within these pathways is often associated withvarious cancers. Thus, compounds which inhibit the action of PRMTs, asprovided herein, are effective in the treatment of cancer.

In some embodiments, compounds provided herein are effective in treatingcancer through the inhibition of PRMT1. For example, PRMT1overexpression has been observed in various human cancers, including,but not limited to, breast cancer, prostate cancer, lung cancer, coloncancer, bladder cancer, and leukemia. In one example, PRMT1 specificallydeposits an asymmetric dimethylarginine (aDMA) mark on histone H4 atarginine 3 (H4R3me2a), and this mark is associated with transcriptionactivation. In prostate cancer, the methylation status of H4R3positively correlates with increasing tumor grade and can be used topredict the risk of prostate cancer recurrence (Seligson et al., Nature2005 435, 1262-1266). Thus, in some embodiments, inhibitors of PRMT1, asdescribed herein, are useful in treating cancers associated with themethylation status of H4R3, e.g., prostate cancer. Additionally, themethylarginine effector molecule TDRD3 interacts with the H4R3me2a mark,and overexpression of TDRD3 is linked to poor prognosis for the survivalof patients with breast cancer (Nagahata et al., Cancer Sci. 2004 95,218-225). Thus, in some embodiments, inhibitors of PRMT1, as describedherein, are useful in treating cancers associated with overexpression ofTDRD3, e.g., breast cancer, as inhibition of PRMT1 leads to a decreasein methylation of H4R3, thereby preventing the association ofoverexpressed TDRD3 with H4R3me2a. In other examples, PRMT1 is known tohave non-histone substrates. For example, PRMT1, when localized to thecytoplasm, methylates proteins that are involved in signal transductionpathways, e.g., the estrogen receptor (ER). The expression status of ERin breast cancer is critical for prognosis of the disease, and bothgenomic and non-genomic ER pathways have been implicated in thepathogenesis of breast cancer. For example, it has been shown that PRMT1methylates ERα, and that ERα methylation is required for the assembly ofERα with SRC (a proto-oncogene tyrosine-protein kinase) and focaladhesion kinase (FAK). Further, the silencing of endogenous PRMT1resulted in the inability of estrogen to activate AKT. These resultssuggested that PRMT1-mediated ERα methylation is required for theactivation of the SRC-PI3K-FAK cascade and AKT, coordinating cellproliferation and survival. Thus, hypermethylation of ERα in breastcancer is thought to cause hyperactivation of this signaling pathway,providing a selective survival advantage to tumor cells (Le Romancer etal., Mol. Cell 2008 31, 212-221; Le Romancer et al., Steroids 2010 75,560-564). Accordingly, in some embodiments, inhibitors of PRMT1, asdescribed herein, are useful in treating cancers associated with ERαmethylation, e.g., breast cancer. In yet another example, PRMT1 has beenshown to be involved in the regulation of leukemia development. Forexample, SRC-associated in mitosis 68 kDa protein (SAM68; also known asKHDRBS1) is a well-characterized PRMT1 substrate, and when either SAM68or PRMT1 is fused directly to the myeloid/lymphoid leukemia (MLL) gene,these fusion proteins can activate MLL oncogenic properties, implyingthat the methylation of SAM68 by PRMT1 is a critical signal for thedevelopment of leukemia (Cheung et al., Nature Cell Biol. 2007 9,1208-1215). Accordingly, in some embodiments, inhibitors of PRMT1, asdescribed herein, are useful in treating cancers associated with SAM68methylation, e.g., leukemia. In still another example, PRMT1 isimplicated in leukemia development through its interaction with AE9a, asplice isoform of AML1-ETO (Shia et al., Blood 2012 119:4953-62).Knockdown of PRMT1 affects expression of certain AE9a-activated genesand suppresses AE9a's self-renewal capability. It has also been shownthat AE9a recruits PRMT1 to AE9a activated gene promoters, which leadsto increased H4 Arg3 methylation, H3 Lys9/14 acetylation, andtranscription activated. Accordingly, in some embodiments, inhibitors ofPRMT1, as described herein, are useful in treating cancers associatedwith AML1-ETO, e.g., leukemia. Thus, without being bound by anyparticular mechanism, the inhibition of PRMT1, e.g., by compoundsdescribed herein, is beneficial in the treatment of cancer.

In some embodiments, compounds provided herein are effective in treatingcancer through the inhibition of PRMT3. In one example, the DAL1 tumorsuppressor protein has been shown to interact with PRMT3 and inhibitsits methyltransferase activity (Singh et al., Oncogene 2004 23,7761-7771). Epigenetic downregulation of DAL1 has been reported inseveral cancers (e.g., meningiomas and breast cancer), thus PRMT3 isexpected to display increased activity, and cancers that display DAL1silencing may, in some aspects, be good targets for PRMT3 inhibitors,e.g., those described herein. Thus, without being bound by anyparticular mechanism, the inhibition of PRMT3, e.g., by compoundsdescribed herein, is beneficial in the treatment of cancer.

In some embodiments, compounds provided herein are effective in treatingcancer through the inhibition of PRMT4, also known as CARM1. Forexample, PRMT4 levels have been shown to be elevated incastration-resistant prostate cancer (CRPC), as well as in aggressivebreast tumors (Hong et al., Cancer 2004 101, 83-89; Majumder et al.,Prostate 2006 66, 1292-1301). Thus, in some embodiments, inhibitors ofPRMT4, as described herein, are useful in treating cancers associatedwith PRMT4 overexpression. PRMT4 has also been shown to affectERα-dependent breast cancer cell differentiation and proliferation(Al-Dhaheri et al., Cancer Res. 2011 71, 2118-2128), thus in someaspects PRMT4 inhibitors, as described herein, are useful in treatingERα-dependent breast cancer by inhibiting cell differentiation andproliferation. In another example, PRMT4 has been shown to be recruitedto the promoter of E2F1 (which encodes a cell cycle regulator) as atranscriptional co-activator (Frietze et al., Cancer Res. 2008 68,301-306). Thus, PRMT4-mediated upregulation of E2F1 expression maycontribute to cancer progression and chemoresistance as increasedabundance of E2F1 triggers invasion and metastasis by activating growthreceptor signaling pathways, which in turn promote an antiapoptotictumor environment (Engelmann and Pützer, Cancer Res 2012 72; 571).Accordingly, in some embodiments, the inhibition of PRMT4, e.g., bycompounds provided herein, is useful in treating cancers associated withE2F1 upregulation. Thus, without being bound by any particularmechanism, the inhibition of PRMT4, e.g., by compounds described herein,is beneficial in the treatment of cancer.

In some embodiments, compounds provided herein are effective in treatingcancer through the inhibition of PRMT6. For example, PRMT6 has beenreported to be overexpressed in a number of cancers, e.g., bladder andlung cancer (Yoshimatsu et al., Int. J. Cancer 2011 128, 562-573). Thus,in some embodiments, the inhibition of PRMT6, by compounds providedherein, is useful in treating cancers associated with PRMT6overexpression. In some aspects, PRMT6 is primarily thought to functionas a transcriptional repressor, although it has also been reported thatPRMT6 functions as a co-activator of nuclear receptors. For example, asa transcriptional repressor, PRMT6 suppresses the expression ofthrombospondin 1 (TSP1; also known as THBS1; a potent natural inhibitorof angiogenesis and endothelial cell migration) and p21 (a naturalinhibitor of cyclin dependent kinase), thereby contributing to cancerdevelopment and progression (Michaud-Levesque and Richard, J Biol. Chem.2009 284, 21338-21346; Kleinschmidt et al., PLoS ONE 2012 7, e41446).Accordingly, in some embodiments, the inhibition of PRMT6, by compoundsprovided herein, is useful in treating cancer by preventing therepression of THBs1 and/or p21. Thus, without being bound by anyparticular mechanism, the inhibition of PRMT6, e.g., by compoundsdescribed herein, is beneficial in the treatment of cancer.

In some embodiments, compounds provided herein are effective in treatingcancer through the inhibition of PRMT8. For example, deep-sequencingefforts of cancer genomes (e.g., COSMIC) have revealed that of all thePRMTs, PRMT8 is reported to be the most mutated. Of 106 sequencedgenomes, 15 carry mutations in the PRMT8 coding region, and nine ofthese result in an amino acid change (Forbes et al., Nucleic Acids Res.2011 39, D945-D950). Because of its high rate of mutation in cancer,PRMT8 is thought to contribute to the initiation or progression ofcancer. Thus, without being bound by any particular mechanism, theinhibition of PRMT8, e.g., by compounds described herein, is beneficialin the treatment of cancer.

In some embodiments, compounds described herein are useful for treatinga cancer including, but not limited to, acoustic neuroma,adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g.,lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma),appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g.,cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinomaof the breast, papillary carcinoma of the breast, mammary cancer,medullary carcinoma of the breast), brain cancer (e.g., meningioma;glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchuscancer, carcinoid tumor, cervical cancer (e.g., cervicaladenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma,colorectal cancer (e.g., colon cancer, rectal cancer, colorectaladenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma(e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma),endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophagealcancer (e.g., adenocarcinoma of the esophagus, Barrett's adenocarinoma),Ewing sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma),familiar hypereosinophilia, gall bladder cancer, gastric cancer (e.g.,stomach adenocarcinoma), gastrointestinal stromal tumor (GIST), head andneck cancer (e.g., head and neck squamous cell carcinoma, oral cancer(e.g., oral squamous cell carcinoma (OSCC), throat cancer (e.g.,laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer,oropharyngeal cancer)), hematopoietic cancers (e.g., leukemia such asacute lymphocytic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acutemyelocytic leukemia (AML) (e.g., B-cell AML, T-cell AML), chronicmyelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML), and chroniclymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL); lymphoma suchas Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkinlymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma(DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicularlymphoma, chronic lymphocytic leukemia/small lymphocytic lymphoma(CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphomas(e.g., mucosa-associated lymphoid tissue (MALT) lymphomas, nodalmarginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma),primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacyticlymphoma (e.g., “Waldenström's macroglobulinemia”), hairy cell leukemia(HCL), immunoblastic large cell lymphoma, precursor B-lymphoblasticlymphoma and primary central nervous system (CNS) lymphoma; and T-cellNHL such as precursor T-lymphoblastic lymphoma/leukemia, peripheralT-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g.,mycosis fungiodes, Sezary syndrome), angioimmunoblastic T-cell lymphoma,extranodal natural killer T-cell lymphoma, enteropathy type T-celllymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplasticlarge cell lymphoma); a mixture of one or more leukemia/lymphoma asdescribed above; and multiple myeloma (MM)), heavy chain disease (e.g.,alpha chain disease, gamma chain disease, mu chain disease),hemangioblastoma, inflammatory myofibroblastic tumors, immunocyticamyloidosis, kidney cancer (e.g., nephroblastoma a.k.a. Wilms' tumor,renal cell carcinoma), liver cancer (e.g., hepatocellular cancer (HCC),malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, smallcell lung cancer (SCLC), non-small cell lung cancer (NSCLC),adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis (e.g.,systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma,myeloproliferative disorder (MPD) (e.g., polycythemia Vera (PV),essential thrombocytosis (ET), agnogenic myeloid metaplasia (AMM) a.k.a.myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocyticleukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilicsyndrome (HES)), neuroblastoma, neurofibroma (e.g., neurofibromatosis(NF) type 1 or type 2, schwannomatosis), neuroendocrine cancer (e.g.,gastroenteropancreatic neuroendoctrine tumor (GEP-NET), carcinoidtumor), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarianembryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma,pancreatic cancer (e.g., pancreatic andenocarcinoma, intraductalpapillary mucinous neoplasm (IPMN), Islet cell tumors), penile cancer(e.g., Paget's disease of the penis and scrotum), pinealoma, primitiveneuroectodermal tumor (PNT), prostate cancer (e.g., prostateadenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary gland cancer,skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA),melanoma, basal cell carcinoma (BCC)), small bowel cancer (e.g.,appendix cancer), soft tissue sarcoma (e.g., malignant fibroushistiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor(MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous glandcarcinoma, sweat gland carcinoma, synovioma, testicular cancer (e.g.,seminoma, testicular embryonal carcinoma), thyroid cancer (e.g.,papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC),medullary thyroid cancer), urethral cancer, vaginal cancer and vulvarcancer (e.g., Paget's disease of the vulva).

In some embodiments, a compound provided herein is useful in treatingdiseases associated with increased levels of circulating asymmetricdimethylarginine (aDMA), e.g., cardiovascular disease, diabetes, kidneyfailure, renal disease, pulmonary disease, etc. Circulating aDMA isproduced by the proteolysis of asymmetrically dimethylated proteins.PRMTs which mediate aDMA methylation include, e.g., PRMT1, PRMT3, PRMT4,PRMT6, and PRMT8. aDMA levels are directly involved in various diseasesas aDMA is an endogenous competitive inhibitor of nitric oxide synthase(NOS), thereby reducing the production of nitric oxide (NO) (Vallance etal., J. Cardiovasc. Pharmacol. 1992 20(Suppl. 12):S60-2). NO functionsas a potent vasodilator in endothelial vessels, and as such inhibitingits production has major consequences on the cardiovascular system. Forexample, since PRMT1 is a major enzyme that generates aDMA, thedysregulation of its activity is likely to regulate cardiovasculardiseases (Boger et al., Ann. Med. 2006 38:126-36), and otherpathophysiological conditions such as diabetes mellitus (Sydow et al.,Vasc. Med. 2005 10(Suppl. 1):S35-43), kidney failure (Vallance et al.,Lancet 1992 339:572-5), and chronic pulmonary diseases (Zakrzewicz etal., BMC Pulm. Med. 2009 9:5). Additionally, it has been demonstratedthat the expression of PRMT1 and PRMT3 are increased in coronary heartdisease (Chen et al., Basic Res. Cardiol. 2006 101:346-53). In anotherexample, aDMA elevation is seen in patients with renal failure, due toimpaired clearance of this metabolite from the circulation (Jacobi etal., Am. J. Nephrol. 2008 28:224-37). Thus, circulating aDMA levels isobserved in many pathophysiological situations. Accordingly, withoutbeing bound by any particular mechanism, the inhibition of PRMTs, e.g.,by compounds described herein, results in the decrease of circulatingaDMA, which is beneficial in the treatment of diseases associated withincreased levels of circulating aDMA, e.g., cardiovascular disease,diabetes, kidney failure, renal disease, pulmonary disease, etc. Incertain embodiments, a compound described herein is useful for treatingor preventing vascular diseases.

In some embodiments, a compound provided herein is useful in treatingmetabolic disorders. For example, PRMT1 has been shown to enhance mRNAlevels of FoxO1 target genes in gluconeogenesis, which results inincreased hepatic glucose production, and knockdown of PRMT promotesinhibition of FoxO1 activity and thus inhibition of hepaticgluconeogenesis (Choi et al., Hepatology 2012 56:1546-56). Additionally,genetic haploinsufficiency of Prmt1 has been shown to reduce bloodglucose levels in mouse models. Thus, without being bound by anyparticular mechanism, the inhibition of PRMT1, e.g., by compoundsdescribed herein, is beneficial in the treating of metabolic disorders,such as diabetes. In some embodiments, a provided compound is useful intreating type I diabetes. In some embodiments, a provided compound isuseful in treating type II diabetes.

In some embodiments, a compound provided herein is useful in treatingmuscular dystrophies. For example, PRMT1, as well as PRMT3 and PRMT6,methylate the nuclear poly(A)-binding protein (PABPN1) in a regionlocated near its C-terminus (Perreault et al., J. Biol. Chem. 2007282:7552-62). This domain is involved in the aggregation of the PABPN1protein, and abnormal aggregation of this protein is involved in thedisease oculopharyngeal muscular dystrophy (Davies et al., Int. J.Biochem. Cell. Biol. 2006 38:1457-62). Thus, without being bound by anyparticular mechanism, the inhibition of PRMTs, e.g., by compoundsdescribed herein, is beneficial in the treatment of musculardystrophies, e.g., oculopharyngeal muscular dystrophy, by decreasing theamount of methylation of PABPN1, thereby decreasing the amount of PABPN1aggregation.

CARM1 is also the most abundant PRMT expressed in skeletal muscle cells,and has been found to selectively control the pathways modulatingglycogen metabolism, and associated AMPK (AMP-activated protein kinase)and p38 MAPK (mitogen-activated protein kinase) expression. See, e.g.,Wang et al., Biochem (2012) 444:323-331. Thus, in some embodiments,inhibitors of CARM1, as described herein, are useful in treatingmetabolic disorders, e.g., for example skeletal muscle metabolicdisorders, e.g., glycogen and glucose metabolic disorders. Exemplaryskeletal muscle metabolic disorders include, but are not limited to,Acid Maltase Deficiency (Glycogenosis type 2; Pompe disease), Debrancherdeficiency (Glycogenosis type 3), Phosphorylase deficiency (McArdle's;GSD 5), X-linked syndrome (GSD9D), Autosomal recessive syndrome (GSD9B),Tarui's disease (Glycogen storage disease VII; GSD 7), PhosphoglycerateMutase deficiency (Glycogen storage disease X; GSDX; GSD 10), Lactatedehydrogenase A deficiency (GSD 11), Branching enzyme deficiency (GSD4), Aldolase A (muscle) deficiency, β-Enolase deficiency,Triosephosphate isomerase (TIM) deficiency, Lafora's disease(Progressive myoclonic epilepsy 2), Glycogen storage disease (Muscle,Type 0, Phosphoglucomutase 1 Deficiency (GSD 14)), and GlycogeninDeficiency (GSD 15).

In some embodiments, a compound provided herein is useful in treatingautoimmune disease. For example, several lines of evidence stronglysuggest that PRMT inhibitors may be valuable for the treatment ofautoimmune diseases, e.g., rheumatoid arthritis. PRMTs are known tomodify and regulate several critical immunomodulatory proteins. Forexample, post-translational modifications (e.g., arginine methylation),within T cell receptor signaling cascades allow T lymphocytes toinitiate a rapid and appropriate immune response to pathogens.Co-engagement of the CD28 costimulatory receptor with the T cellreceptor elevates PRMT activity and cellular protein argininemethylation, including methylation of the guanine nucleotide exchangefactor Vav1 (Blanchet et al., J. Exp. Med. 2005 202:371-377). PRMTinhibitors are thus expected to diminish methylation of the guanineexchange factor Vav1, resulting in diminished IL-2 production. Inagreement, siRNA directed against PRMT5 was shown to both inhibitNFAT-driven promoter activity and IL-2 secretion (Richard et al.,Biochem J. 2005 388:379-386). In another example, PRMT1 is known tocooperate with PRMT4 to enhance NFkB p65-driven transcription andfacilitate the transcription of p65 target genes like TNFα (Covic etal., Embo. J. 2005 24:85-96). Thus, in some embodiments, PRMT1 and/orPRMT4 inhibitors, e.g., those described herein, are useful in treatingautoimmune disease by decreasing the transcription of p65 target geneslike TNFα. These examples demonstrate an important role for argininemethylation in inflammation. Thus, without being bound by any particularmechanism, the inhibition of PRMTs, e.g., by compounds described herein,is beneficial in the treatment of autoimmune diseases.

In some embodiments, a compound provided herein is useful in treatingneurological disorders, such as amyotrophic lateral sclerosis (ALS). Forexample, a gene involved in ALS, TLS/FUS, often contains mutatedarginines in certain familial forms of this disease (Kwiatkowski et al.,Science 2009 323:1205-8). These mutants are retained in the cytoplasm,which is similar to reports documenting the role arginine methylationplays in nuclear-cytoplasmic shuffling (Shen et al., Genes Dev. 199812:679-91). This implicates PRMT, e.g., PRMT1, function in this disease,as it was demonstrated that TLS/FUS is methylated on at least 20arginine residues (Rappsilber et al., Anal. Chem. 2003 75:3107-14).Thus, in some embodiments, the inhibition of PRMTs, e.g., by compoundsprovided herein, are useful in treating ALS by decreasing the amount ofTLS/FUS arginine methylation.

Scheme 1 shows an exemplary general synthesis route to pyrazolecompounds of formula I, wherein R^(W′) is either the same as R^(W) or isprecursor of R^(W) and L_(1′) is either the same as L₁ or is a precursorof L₁ and R^(W), L₁, R^(x), R³, X, Y and Z are as defined above. In thefirst step iodopyrazole carboxaldehydes of general formula XI areallowed to react with mono-Boc protected ethylenediamines XII underreductive amination conditions (e.g. sodium cyanoborohydride andcatalytic acid such as acetic acid) in an appropriate solvent such asmethanol to give intermediates of general formula XIII. In certainembodiments, Sonagashira reaction of intermediates of general formulaXIII with boronic acids or boronic esters of general formula XIV inwhich L_(1′) is an acetylene linker and Q is a boronic acid or boronicester group in the presence of a palladium catalyst (e.g. PdCl₂(dppf))and a base (e.g. potassium carbonate) in an organic solvent (e.g.toluene) at elevated temperature yields intermediates of general formulaXV-a in which L_(1′) is an acetylene linker. Boc deprotection ofintermediates of general formula XV-a gives acetylene compounds offormula VI-a. In certain embodiments, Suzuki reaction of intermediatesof general formula XIII with boronic acids or boronic esters of generalformula XIV in which L_(1′) is a trans-olefin linker and Q is a boronicacid or boronic ester group in the presence of a palladium catalyst(e.g. PdCl₂(dppf)) and a base (e.g. potassium carbonate) in an organicsolvent (e.g. toluene) at elevated temperature yields intermediates ofgeneral formula XV-b in which L_(1′) is an olefin linker. Bocdeprotection of intermediates of general formula XV-b gives olefincompounds of formula VI-b. In certain embodiments, Suzuki reaction ofintermediates of general formula XIII with pinacol boranes of generalformula XIVc in which L_(1′) is bond, R^(W′) is a heterocycloalkenyl orcycloalkenyl group and Q is a pinacol borane group yields intermediatesof general formula XV-c in which L_(1′) is bond and R^(W′) is aheterocycloalkenyl or cycloalkenyl group. In certain embodiments,compounds of formula I wherein L₁ is bond and R^(W) is a heterocyclyl orcarbocyclyl group can be prepared by hydrogenation of intermediates offormula XV-c followed by Boc deprotection. In certain embodiments,compounds of formula I where L₁ is —O— can be synthesized fromintermediates of general formula XIII by Goldberg reaction with alcoholsof formula R^(W)OH followed by Boc deprotection. In certain embodiments,compounds of formula I where L₁ is —N(R^(B))— can be synthesized fromintermediates of general formula XIII by palladium catalyzed Buchwaldcoupling reaction conditions with amines of formula R^(W)N(R^(B))Hfollowed by Boc deprotection. In certain embodiments, compounds offormula I where L₁ is —C(═O)NR^(B)— can be synthesized fromintermediates of general formula XIII under known copper catalyzedcoupling reaction conditions of amides with aryliodides using copperiodide an amine ligand and a base with amides of formulaR^(W)C(═O)NHR^(B) followed by Boc deprotection.

Scheme 1.1 shows an alternative general synthesis route to pyrazolecompounds of Formula (I), that involves reversal in the order of thefirst two steps of the reaction sequence detailed for Scheme 1.0. Thus,in the first step iodopyrazole carboxaldehydes of general formula XI arecoupled with compounds or reagents of general formula XIV (e.g. viaSuzuki reaction with pinacol boranes of general formula XIVc in whichL_(1′) is bond, R^(W′) is a heterocycloalkenyl or cycloalkenyl group andQ is a pinacol borane group) and in a second step the correspondingreductive amination reaction to yield common intermediates of generalformula XV is a carried out.

In certain embodiments, iodopyrazole carboxaldehydes of general formulaXI may be prepared from suitable known pyrazole compound intermediatesby established synthetic chemistry methods. Standard methods includedirect iodination of a pyrazole 3-carboxylate and Sandmeyer reaction ofa 3-amino pyrazole 4-carboxylate. In certain embodiments, iodopyrazolecarboxaldehydes can be derived from iodopyrazole carboxylates byreduction to a hydroxymethyl group followed by oxidation tocarboxaldehyde. In certain embodiments, mono-Boc protectedethylenediamines XII can be synthesized by standard methods known in theliterature for derivatizing or preparing ethylenediamines. For exampleintermediates of formula XII may be prepared by treatment of thecorresponding unprotected diamine precursors with Boc₂O and purifyingthe mixture of mono and dibocylated products. In certain embodiments,pyrazole compounds of general formula II can be prepared fromiodopyrazole carboxaldehydes of general formula XXI as depicted inScheme 2. In certain embodiments where R⁴ is hydrogen compounds ofgeneral formula II are equivalent to compounds of general formula IIIwhich are tautomers. In certain embodiments, R^(4′) is a protectinggroup such as tetrahydropyranyl (THP) which maybe cleaved to hydrogenunder acidic conditions in the final Boc-deprotection step. In certainembodiments, iodopyrazole carboxaldehydes of general formula XXI can beprepared as depicted in Scheme 3.

In certain embodiments, iodopyrazole carboxaldehydes of general formulaXXI can be prepared as depicted in Scheme 4 which also providesiodopyrazole carboxyaldehydes of general formula XXXI. In certainembodiments, alkylation of intermediates of general formula XXX gives amixture of pyrazole nitrogen alkylated isomers which are separated bychromatography to give pure isomers XXI and XXXI. In certainembodiments, pyrazole compounds of general formula III can be preparedfrom iodopyrazole carboxaldehydes of general formula XXXI as depicted inScheme 5.

In certain embodiments, pyrazole compounds of general formula IV can beprepared from iodopyrazole carboxaldehydes of general formula XLI asdepicted in Scheme 6. In certain embodiments where R⁴ is hydrogencompounds of general formula IV are equivalent to compounds of generalformula V which are tautomers. In certain embodiments where R⁴ incompounds of formula IV is hydrogen, R^(4′) in intermediate XLI may be aselected protecting group such as tetrahydropyranyl (THP) which can becleaved to hydrogen under acidic conditions in the finalBoc-deprotection step.

In certain embodiments, iodopyrazole carboxaldehydes of general formulaXLI and LI can be prepared as depicted in Scheme 7. In certainembodiments, an R⁴ group of iodopyrazole carboxaldehydes may beintroduced by alkylation of intermediates of formula XLVII. Thisreaction can give a mixture of intermediate compounds of formulas XLIand LI which may be separated by chromatography. In certain embodiments,THP protected intermediates of formula XLVI can be used to preparecompounds of formula IV where R⁴═H as also depicted in Scheme 7.

In certain embodiments, pyrazole compounds of general formula V can beprepared from iodopyrazole carboxaldehydes of general formula LI asdepicted in Scheme 8.

In certain embodiments, boronic acids or esters of general formula XIVa,XIVb and XIVc are commercially available. In certain embodiments,compounds of general formula XIVa, and XIVb can also be prepared fromalkenyl bromides and terminal alkynes using standard methods such astreatment with n-BuLi followed by trapping the intermediate lithiumspecies with trimethylborate. In certain embodiments, compounds ofgeneral formula XIVc can be prepared from the corresponding cyclicketones LX via intermediate enol triflates as depicted in Scheme 9.

EXAMPLES

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

Synthetic Methods

General methods and experimental procedures for preparing andcharacterizing compounds of the present invention are set forth below.Wherever needed, reactions were heated using conventional hotplateapparatus or heating mantle or microwave irradiation equipment.Reactions were conducted with or without stirring, under atmospheric orelevated pressure in either open or closed vessels. Reaction progresswas monitored using conventional techniques such as TLC, HPLC, UPLC, orLCMS using instrumentation and methods described below. Reactions werequenched and crude compounds isolated using conventional methods asdescribed in the specific examples provided. Solvent removal was carriedout with or without heating, under atmospheric or reduced pressure,using either a rotary or centrifugal evaporator.

Compound purification was carried out as needed using a variety oftraditional methods including, but not limited to, preparativechromatography under acidic, neutral, or basic conditions using eithernormal phase or reverse phase HPLC or flash columns or Prep-TLC plates.Compound purity and mass confirmations were conducted using standardHPLC and/or UPLC and/or MS spectrometers and/or LCMS and/or GC equipment(e.g., including, but not limited to the following instrumentation:Waters Alliance 2695 with 2996 PDA detector connected with ZQ detectorand ESI source; Shimadzu LDMS-2020; Waters Acquity H Class with PDAdetector connected with SQ detector and ESI source; Agilent 1100 Serieswith PDA detector; Waters Alliance 2695 with 2998 PDA detector; AB SCIEXAPI 2000 with ESI source; Agilent 7890 GC). Exemplified compounds weredissolved in either MeOH or MeCN to a concentration of approximately 1mg/mL and analyzed by injection of 0.5-10 μL into an appropriate LCMSsystem using the methods provided in the following table:

MS Heat MS Flow Block Detector Mobile Mobile Rate Temp Voltage MethodColumn Phase A Phase B (mL/min) Gradient Profile (° C.) (kV) A Shim-packWater/0.05% ACN/0.05% 1 5% to 100% B in 2.0 250 1.5 XR-ODS TFA TFAminutes, 100% B for 2.2 μm 1.1 minutes, 100% to 3.0 × 50 mm 5% B in 0.2minutes, then stop B Gemini-NX Water/ ACN 1 5% to 100% B in 2.0 200 0.753 μm C18 0.04% minutes, 100% B for 110A Ammonia 1.1 minutes, 100% to 5%B in 0.1 minutes, then stop C Shim-pack Water/0.05% ACN/0.05% 1 5% to100% B in 2.0 250 0.85 XR-ODS FA FA minutes, 100% B for 1.6 μm 1.1minutes, 100% to 2.0 × 50 mm 5% B in 0.1 minutes, then stop D Shim-packWater/0.05% ACN/0.05% 1 5% to 100% B in 2.0 250 0.95 XR-ODS TFA TFAminutes, 100% B for 2.2 μm 1.1 minutes, 100% to 3.0 × 50 mm 5% B in 0.1minutes, then stop E Waters Water/0.05% ACN/0.05% 0.9 5% to 100% B in2.0 250 1.5 Xselect C18 FA FA minutes, 100% B for 3.5 μm 1.2 minutes,100% to 3.0 × 50 mm 5% B in 0.1 minutes, then stop F Shim-packWater/0.05% ACN/0.05% 1 5% to 80% B in 3.25 200 0.95 XR-ODS TFA TFAminutes, 80% B for 2.2 μm 1.35 minutes, 80% to 3.0 × 50 mm 5% B in 0.3minutes, then stop G Shim-pack Water/0.05% ACN/0.05% 1 5% to 70% B in2.50 200 0.95 XR-ODS TFA TFA minutes, 70% B for 2.2 μm 0.70 minutes, 70%to 3.0 × 50 mm 5% B in 0.1 minutes, then stop H Shim-pack Water/0.05%ACN/0.05% 1 5% to 100% B in 2.20 250 0.95 XR-ODS TFA TFA minutes, 100% Bfor 2.2 μm 1.00 minutes, 100% to 3.0 × 50 mm 5% B in 0.1 minutes, thenstop I Shim-pack Water/0.05% ACN/0.05% 1 5% to 100% B in 1.20 250 0.95XR-ODS TFA TFA minutes, 100% B for 2.2 μm 1.00 minutes, 100% to 3.0 × 50mm 5% B in 0.1 minutes, then stop J Shim-pack Water/0.05% ACN/0.05% 1 5%to 70% B in 3.20 250 0.95 XR-ODS TFA TFA minutes, 70% B for 2.2 μm 0.75minutes, 70% to 3.0 × 50 mm 5% B in 0.35 minutes, then stop K Shim-packWater/0.05% ACN/0.05% 1 5% to 80% B in 3.00 250 1.5 XR-ODS TFA TFAminutes, 80% B for 2.2 μm 0.8 minutes, 80% to 3.0 × 50 mm 5% B in 0.1minutes, then stop L Shim-pack Water/0.05% ACN/0.05% 1 5% to 100% B in3.00 250 1.5 XR-ODS TFA TFA minutes, 100% B for 2.2 μm 0.8 minutes, 100%to 3.0 × 50 mm 5% B in 0.1 minutes, then stop M Shim-pack Water/0.05%ACN/0.05% 1 5% to 100% B in 2.20 250 1.5 XR-ODS TFA TFA minutes, 100% Bfor 2.2 μm 1.00 minutes, 100% to 3.0 × 50 mm 5% B in 0.1 minutes, thenstop N Shim-pack Water/0.05% ACN/0.05% 1 5% to 80% B in 2.20 250 1.5XR-ODS TFA TFA minutes, 80% B for 2.2 μm 1.0 minutes, 80% to 3.0 × 50 mm5% B in 0.1 minutes, then stop O Zorbax Water/0.05% ACN/0.05% 1 5% to70% B in 8.00 250 1.5 Eclipse Plus TFA TFA minutes, 70% B for C18 2.0minutes, then stop 4.6 × 100 mm P Shim-pack Water/0.05% ACN/0.05% 1 5%to 65% B in 3.00 250 1.5 XR-ODS TFA TFA minutes, 65% B for 2.2 μm 0.80minutes, 100% to 3.0 × 50 mm 5% B in 0.1 minutes, then stop Q Shim-packWater/0.05% ACN/0.05% 1 5% to 60% B in 2.50 250 0.95 XR-ODS TFA TFAminutes, 60% B for 2.2 μm 0.7 minutes, 60% to 3.0 × 50 mm 5% B in 0.1minutes, then stop R Shim-pack Water/0.05% ACN/0.05% 1 5% to 50% B in2.50 250 0.95 XR-ODS TFA TFA minutes, 50% B for 2.2 μm 0.7 minutes, 50%to 3.0 × 50 mm 5% B in 0.1 minutes, then stop S XBridge Water/0.05%ACN/0.05% 1 5% to 95% B in 2.20 250 0.9 C18 3.5 μm TFA TFA minutes, 95%B for 3.0 × 50 mm 1.00 minutes, 95% to 5% B in 0.1 minutes, then stop TShim-pack Water/0.05% ACN/0.05% 0.7 5% to 100% B in 2.0 250 0.85 XR-ODSFA FA minutes, 100% B for 1.6 μm 1.1 minutes, 100% to 2.0 × 50 mm 5% Bin 0.1 minutes, then stop U Shim-pack Water/0.05% ACN/0.05% 1 5% to 40%B in 2.50 250 0.95 XR-ODS TFA TFA minutes, 40% B for 2.2 μm 0.7 minutes,40% to 3.0 × 50 mm 5% B in 0.1 minutes, then stop V Shim-packWater/0.05% ACN/0.05% 1 5% to 60% B in 4.20 200 1.05 XR-ODS TFA TFAminutes, 60% B for 2.2 μm 1.0 minutes, 60% to 3.0 × 50 mm 5% B in 0.1minutes, then stop W Shim-pack Water/0.05% ACN/0.05% 1 5% to 100% B in2.20 200 0.95 XR-ODS TFA TFA minutes, 100% B for 2.2 μm 1.00 minutes,100% to 3.0 × 50 mm 5% B in 0.1 minutes, then stop X Shim-packWater/0.05% ACN/0.05% 0.7 5% to 100% B in 2.0 200 0.85 XR-ODS FA FAminutes, 100% B for 1.6 μm 1.1 minutes, 100% to 2.0 × 50 mm 5% B in 0.1minutes, then stop Y Ecliplis Water/0.05% ACN 1 5% to 100% B in 2.0 2501 Plus C18 TFA minutes, 100% B for 3.5 μm 1.0 minutes, 100% to 4.6 × 50mm 5% B in 0.1 minutes, then stop Z Ecliplis Water/10 mM ACN/5% 1 5% to100% B in 2.0 250 1.1 Plus C18 ammonium water minutes, 100% B for 3.5 μmcarbonate 1.0 minutes, 100% to 4.6 × 50 mm 5% B in 0.1 minutes, thenstop A1 Shim-pack Water/0.05% ACN 1 5% to 100% B in 2.0 250 1 XR-ODS TFAminutes, 100% B for 2.2 μm 1.0 minutes, 100% to 3.0 × 50 mm 5% B in 0.1minutes, then stop A2 Ecliplis Water/10 mM ACN 1 5% to 100% B in 2.0 2500.95 Plus C18 ammonium minutes, 100% B for 3.5 μm acetate 1.4 minutes,100% to 4.6 × 50 mm 5% B in 0.1 minutes, then stop A3 Acquity Water/5 mMACN/0.1% 0.55 5% B at 0.01 min up BEH C18 ammonium FA to 0.4 min, 35% Bat 1.7 μm 2.1 × acetate/ 0.8 min, 55% B at 1.2 min, 50 mm 0.1% FA 100% Bin 1.3 minutes, at 2.5 min up to 3.30 min, 5% B at 3.31 min up to 4.0min, then stop A4 Shim-pack Water/0.05% ACN/0.05% 1 5% to 30% B in 8.0250 1.5 XR-ODS TFA TFA minutes, 30% B for 3.0 × 50 mm 2.0 minutes, thenstop A5 Shim-pack Water/0.05% ACN/0.05% 1 5% to 100% B in 2.2 250 1.5XR-ODS TFA TFA minutes, 100% B for 3.0 × 50 mm 1.0 minutes, 100% to 5% Bin 0.1 minutes, then stop A6 Atlantis Water/0.05% ACN/0.05% 0.8 95% to60% B in 4.0 250 1.5 HILIC TFA TFA minutes, 60% B for 3.0 × 100 mm 4.0minutes, then stop A7 Shim-pack Water/0.05% ACN/0.05% 1 5% B for 0.5minutes, 250 1.5 XR-ODS TFA TFA 5% to 75% B at 2.2 3.0 × 50 mm minutes,100% B for 1.0 minutes, 100% to 5% B in 0.1 minutes, then stop A8 ZorbaxSB- Water/0.05% ACN/0.05% 1.2 5% to 70% B in 10.0 250 1.05 C18 TFA TFAminutes, 70% B for 5 μm 5.0 minutes, then stop 4.6 × 150 mm A9 Shim-packWater/0.05% ACN/0.05% 1 5% to 40% B in 4.4 250 0.95 XR-ODS TFA TFAminutes, 40% B for 3.0 × 50 mm 0.9 minutes, then stop A10 Atlantis T3Water/0.05% ACN/0.05% 1 5% to 50% B in 8.0 200 1.05 3 μm TFA TFAminutes, 50% B for 4.6 × 100 mm 2.0 minutes, then stop A11 Shim-packWater/0.05% ACN/0.05% 1 5% B for 0.5 minutes, 250 1.50 XR-ODS TFA TFA 5%to 100% B in 1.7 3.0 × 50 mm minutes, 100% B for 1.0 minute, 100% to 5%B in 0.1 minute, then stop

Compound structure confirmations were carried out using standard 300 or400 MHz NMR spectrometers with NOe's conducted whenever necessary.

The following abbreviations are used herein:

Abbreviation Meaning ACN acetonitrile atm. atmosphere DCMdichloromethane DHP dihydropyran DIBAL diisobutyl aluminum hydride DIEAdiisopropyl ethylamine DMF dimethyl formamide DMF-DMA dimethyl formamidedimethyl acetal DMSO dimethyl sulfoxide dppf1,1′-bis(diphenylphosphino)ferrocene EA ethyl acetate ESI electrosprayionization EtOH ethanol FA formic acid GC gas chromatography h hour Hexhexanes HMDS hexamethyl disilazide HPLC high performance liquidchromatography IPA isopropanol LCMS liquid chromatography/massspectrometry MeOH methanol min minutes NBS N-bromo succinimide NCSN-chloro succinimide NIS N-iodo succinimide NMR nuclear magneticresonance NOe nuclear Overhauser effect Prep. preparative PTSApara-toluene sulfonic acid Rf retardation factor rt room temperature RTretention time sat. saturated SGC silica gel chromatography TBAFtetrabutyl ammonium fluoride TEA triethylamine TFA trifluoroacetic acidTHF tetrahydrofuran TLC thin layer chromatography UPLC ultra performanceliquid chromatography LiHMDS lithium hexamethyldisilazide TMADtetramethyl azocarboxamide

Intermediate Synthesis Synthesis of Intermediate tert-butyl(2-(((3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate

Step 1: tert-butyl(2-(((3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate

A mixture of 3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde (3.2 g,10.45 mmol, 1.00 equiv), tert-butyl N-[2-(methylamino)ethyl]carbamate(2.2 g, 12.63 mmol, 1.21 equiv) and NaBH(OAc)₃ (6.65 g, 31.38 mmol, 3.00equiv) in dichloroethane (30 mL) was stirred for 2 h at roomtemperature. The reaction was quenched with 50 mL of saturated aqueoussodium bicarbonate solution. The resulting mixture was extracted with3×200 mL of dichloromethane. The combined organic layers was dried overanhydrous sodium sulfate and concentrated under vacuum. The residue waspurified on a silica gel column eluted with 30-100% ethyl acetate inpetroleum ether to give 4.05 g (83%) of tert-butyl(2-(((3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamateas a light yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.48 (s, 1H),5.35-5.30 (m, 1H), 4.13-4.03 (m, 1H), 3.71-3.63 (m, 1H), 3.36 (s, 2H),3.26-3.25 (m, 2H), 2.52-2.49 (m, 2H), 2.21 (s, 3H), 2.09-2.01 (m, 3H),1.68-1.58 (m, 3H), 1.44 (s, 9H) ppm. LCMS (method C, ESI): RT=0.58 min,m/z=465.0 [M+H]⁺.

Synthesis of Intermediate tert-butyl(2-(((3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)(methyl)carbamate

Step 1: Ethyl 3-iodo-1H-pyrazole-4-carboxylate

To a stirred solution of ethyl 3-amino-1H-pyrazole-4-carboxylate (10 g,64.45 mmol, 1.00 equiv) in 50% sulfuric acid (90 mL) at 5° C. was addeddropwise a solution of NaNO₂ (7.4 g, 107.25 mmol, 1.66 equiv) in water(15 mL). The reaction was stirred at 5° C. for another 30 min. Asolution of KI (32.1 g, 193.37 mmol, 3.00 equiv) in water (15 mL) wasadded dropwise at 5° C. The reaction was allowed to stir at 5° C. for 1h and then quenched by the addition of 50 mL of water. The precipitatewas collected by filtration and then dissolved in 150 mL of ethylacetate. The resulting solution was washed sequentially with 1×100 mL ofsaturated Na₂SO₃ solution, 1×100 mL of saturated sodium bicarbonatesolution and 1×100 mL of brine. The organic layer was dried overanhydrous sodium sulfate and concentrated under vacuum to give 10.8 g(63%) of ethyl 3-iodo-1H-pyrazole-4-carboxylate as a yellow solid.¹H-NMR (300 MHz, CDCl₃): δ 8.18 (s, 1H), 4.38-4.29 (m, 2H), 1.41-1.33(m, 3H) ppm. LCMS (method B, ESI): RT=1.36 min, m/z=267.0 [M+H]⁺.

Step 2: Ethyl3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-carboxylate

A solution of ethyl 3-iodo-1H-pyrazole-4-carboxylate (10.8 g, 40.60mmol, 1.00 equiv), 3,4-dihydro-2H-pyran (10 g, 118.88 mmol, 2.93 equiv)and TsOH (780 mg, 4.53 mmol, 0.11 equiv) in THF (100 mL) was stirred for2 h at 60° C. The reaction mixture was cooled to room temperature andquenched by the addition of 100 mL of saturated sodium bicarbonatesolution. The resulting solution was extracted with 2×80 mL ofdichloromethane. The combined organic layers was dried over anhydroussodium sulfate and concentrated under vacuum. The residue was purifiedon a silica gel column eluted with ethyl acetate/petroleum ether (1:20)to give 13 g (91%) of ethyl3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carboxylate as a yellow oil. ¹H-NMR(400 MHz, CDCl₃): δ 8.04 (s, 1H), 5.40-5.38 (m, 1H), 4.34-4.29 (m, 2H),4.08-4.05 (m, 1H), 3.73-3.70 (m, 1H), 2.07-1.98 (m, 3H), 1.69-1.62 (m,3H), 1.39-1.32 (m, 3H) ppm. LCMS (method C, ESI): RT=1.53 min, m/z=351.0[M+H]⁺.

Step 3: 3-Iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-carboxylicacid

To a solution of ethyl 3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carboxylate(85 g, 242.75 mmol, 1.00 equiv) in THF (300 mL) and methanol (300 mL)was added a solution of LiOH (17.5 g, 730.69 mmol, 3.01 equiv) in water(400 mL). The resulting solution was stirred at room temperatureovernight and then concentrated under vacuum to remove the organicsolvent. The resulting solution was diluted with 400 mL of H₂O and thenacidified to pH 6.0 with 1M hydrochloric acid. The mixture was extractedwith 3×800 mL of dichloromethane. The combined organic layers was washedwith 3×1000 mL of brine, dried over anhydrous sodium sulfate andconcentrated under vacuum to give 75 g (96%) of3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carboxylic acid as an off-whitesolid. LCMS (method D, ESI): RT=1.23 min, m/z=323.0 [M+H]⁺.

Step 4: (3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methanol

To a solution of 3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carboxylic acid (28g, 86.93 mmol, 1.00 equiv) in anhydrous THF (300 mL) maintained undernitrogen at 5° C. was added a 1M solution of BH₃ in THF (300 mL)dropwise with stirring. The reaction was stirred overnight at roomtemperature and then quenched by the addition of 300 mL of saturatedNH₄Cl solution. The resulting mixture was extracted with 3×1000 mL ofdichloromethane. The combined organic layers was dried over anhydroussodium sulfate and concentrated under vacuum. The residue was purifiedon a silica gel column eluted with ethyl acetate/petroleum ether (1:1)to give 12.67 g (47%) of (3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl)methanolas a white solid. ¹H-NMR (400 MHz, DMSO-d6): δ 7.73 (s, 1H), 5.37-5.34(m, 1H), 4.92 (s, 1H), 4.20 (d, J=3.6 Hz, 2H), 3.89-3.88 (m, 1H),3.65-3.57 (m, 1H), 2.09-2.00 (m, 1H), 1.99-1.90 (m, 2H), 1.69-1.61 (m,1H), 1.49-1.46 (m, 2H) ppm. LCMS (method A, ESI): RT=1.16 min, m/z=309.0[M+H]⁺.

Step 5: 3-Iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-carbaldehyde

Into a 250-mL 3-necked round-bottom flask purged and. To a stirredsolution of oxalyl chloride (18.576 g, 146.35 mmol, 3.01 equiv) inanhydrous dichloromethane (300 mL) maintained under nitrogen at −78° C.was added DMSO (15.138 g, 193.75 mmol, 3.98 equiv) dropwise. Thereaction mixture was stirred at −65° C. for 30 min. A solution of(3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl)methanol (15.0 g, 48.68 mmol, 1.00equiv) in dichloromethane (100 mL) was then added dropwise at −65° C.and the reaction was stirred for another 60 min at −65° C. Triethylamine(40.6 mL) was added dropwise at −65° C. and the reaction was stirred for30 min at −65° C. The reaction was warmed to 0° C. then quenched by theaddition of 100 mL of saturated NH₄Cl solution. The resulting mixturewas extracted with 3×400 mL of dichloromethane. The combined organiclayers was dried over anhydrous sodium sulfate and concentrated undervacuum. The residue was purified on a silica gel column eluted withethyl acetate/petroleum ether (1:20) to give 13.48 g (90%) of3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde as a golden oil. ¹H-NMR(300 MHz, DMSO-d6): δ 9.69 (s, 1H), 8.57 (s, 1H), 5.49 (dd, J=2.7 Hz,9.9 Hz, 1H), 3.95-3.91 (m, 1H), 3.68-3.62 (m, 1H), 2.11-2.01 (m, 3H),1.69-1.62 (m, 3H) ppm. LCMS (method A, ESI): RT=1.35 min, m/z=307.0[M+H]⁺.

Step 6: tert-Butyl(2-(((3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)(methyl)carbamate

A mixture of 3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde (21.5 g,70.24 mmol, 1.00 equiv), tert-butylN-methyl-N-(2-(methylamino)ethyl)carbamate (20 g, 106.23 mmol, 1.51equiv) and NaBH(OAc)₃ (29.8 g, 137.98 mmol, 1.96 equiv) indichloroethane (300 mL) was stirred for 1 h at room temperature. Thereaction was diluted with 300 mL of dichloromethane and then washed with3×300 mL of brine. The organic layer was dried over anhydrous sodiumsulfate and concentrated under vacuum. The residue was purified on asilica gel column eluted with 0-7% methanol in dichloromethane to give31 g (92%) of tert-butyl(2-(((3-iodo-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)(methyl)carbamateas a yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.62 (s, 1H), 5.34-5.30 (m,1H), 4.06-4.02 (m, 1H), 3.68-3.62 (m, 1H), 3.42-3.38 (m, 4H), 2.85 (s,4H), 2.62-2.53 (m, 2H), 2.47-2.46 (m, 2H), 2.13-1.97 (m, 3H), 1.74-1.69(m, 3H), 1.46 (s, 9H) ppm. LCMS (method A, ESI): RT=1.17 min, m/z=479.0[M+H]⁺.

Compound 23N¹-((3-(4-fluorophenethyl)-1H-pyrazol-4-yl)methyl)-N′-methylethane-1,2-diamine

Step 1: (R/S)(E)-3-(4-fluorostyryl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-carbaldehyde

A mixture of (R/S) 3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde (800mg, 2.61 mmol, 1.00 equiv), 1-ethenyl-4-fluorobenzene (957 mg, 7.84mmol, 3.00 equiv), Pd(PPh₃)₄(302 mg, 0.26 mmol, 0.10 equiv) andpotassium carbonate (1082 mg, 7.83 mmol, 3.00 equiv) inN,N-dimethylformamide (10 mL) was stirred under nitrogen at 100° C.overnight. The reaction was cooled to room temperature then quenched bythe addition of 100 mL of water. The resulting mixture was extractedwith 3×100 mL of ethyl acetate. The combined organic layers was washedwith 3×100 mL of brine, dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was purified on a silica gelcolumn eluted with 1-15% ethyl acetate in petroleum ether to give 220 mg(28%) of(E)-3-(4-fluorostyryl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-carbaldehydeas a yellow oil. LCMS (method D, ESI): RT=1.49 min, m/z=301.0 [M+H]⁺.

Step 2: (R/S) (E)-tert-butyl2-(((3-(4-fluorostyryl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate

To a solution of (R/S)(E)-3-(4-fluorostyryl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-4-carbaldehyde(220 mg, 0.73 mmol, 1.00 equiv) and tert-butylN-[2-(methylamino)ethyl]carbamate (153 mg, 0.88 mmol, 1.20 equiv) in1,2-dichloroethane (10 mL) was added NaBH(OAc)₃ (311 mg, 1.44 mmol, 1.97equiv). The reaction was stirred at room temperature for 2 h and thendiluted with 100 mL of ethyl acetate. The resulting mixture was washedwith 3×100 mL of brine, dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was purified on a silica gelcolumn eluted with 20-60% ethyl acetate in petroleum ether to give 220mg (65%) of (R/S) (E)-tert-butyl2-(((3-(4-fluorostyryl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamateas a yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 8.20 (br s, 1H), 7.51-7.36(m, 3H), 7.05 (t, J=8.7 Hz, 2H), 6.90 (d, J=15.9 Hz, 1H), 5.38 (t, J=2.7Hz, 1H), 4.12 (s, 2H), 3.75-3.68 (m, 1H), 3.51 (br s, 2H), 2.98 (br s,1H), 2.60 (br s, 2H), 2.19-2.08 (m, 6H), 1.72-1.62 (m, 3H) ppm. LCMS(method D, ESI): RT=1.31 min, m/z=459.2 [M+H]⁺.

Step 3: (R/S) tert-butyl2-(((3-(4-fluorophenethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate

A mixture of (R/S) (E)-tert-butyl2-(((3-(4-fluorostyryl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate(220 mg, 0.48 mmol, 1.00 equiv) and Raney Ni (20 mg) in methanol (50 mL)was stirred under hydrogen at room temperature for 4 h. The catalyst wasremoved by filtration and the filtrate was concentrated under vacuum.The residue was purified on a silica gel column eluted with 1-7% ofethyl acetate in petroleum ether to yield 150 mg (68%) of (R/S)tert-butyl2-(((3-(4-fluorophenethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamateas a colorless oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.11 (t, J=7.8 Hz, 2H),6.94 (t, J=8.7 Hz, 2H), 5.31 (d, J=6.6 Hz, 1H), 4.08 (d, J 11.4 Hz, 1H),3.69 (t, J=11.4 Hz, 1H), 3.44 (br s, 4H), 3.00-2.85 (m, 4H), 2.12-2.09(m, 3H), 1.76-1.52 (m, 6H), 1.45 (s, 9H) ppm. LCMS (method D, ESI):RT=1.29 min, m/z=461.2 [M+H]⁺.

Step 4:N¹-((3-(4-fluorophenethyl)-1H-pyrazol-4-yl)methyl)-N¹-methylethane-1,2-diamine(Compound 23)

A solution of (R/S) tert-butyl2-(((3-(4-fluorophenethyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate(150 mg, 0.33 mmol, 1.00 equiv) in 3N hydrochloric acid (20 mL) wasstirred overnight at 60° C. The resulting mixture was cooled to roomtemperature and washed with 3×20 mL of dichloromethane. The aqueouslayer was concentrated under vacuum and the crude product was purifiedby Prep-HPLC with the following conditions (Prep-HPLC-025): Column,XBridge Prep Phenyl OBD Column, 5 μm, 19×150 mm; mobile phase, waterwith 10 mmol NH₄HCO₃ and MeCN (20.0% MeCN up to 30.0% in 10 min, up to95.0% in 1 min, hold 95.0% in 1 min, down to 20.0% in 2 min); Detector,UV 254/220 nm to give 42.9 mg (26%) ofN¹-((3-(4-fluorophenethyl)-1H-pyrazol-4-yl)methyl)-N¹-methylethane-1,2-diaminetrifluoroacetate as a yellow oil. ¹H-NMR (300 MHz, D₂O) δ: 7.70 (s, 1H),6.98-6.86 (m, 4H), 3.86 (s, 2H), 3.30 (s, 4H), 2.97-2.80 (m, 4H), 2.58(s, 3H) ppm. LCMS (method G, ESI): RT=1.22 min, m/z=277.1 [M+H]⁺.

Compound 28N¹-((3-iso-butyl-1H-pyrazol-4-yl)methyl)-N¹-methylethane-1,2-diamine

Step 1: tert-butyl2-(((3-iso-butyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate

A mixture of (R/S) tert-butylN-[2-([[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl](methyl)amino)ethyl]carbamate(400 mg, 0.86 mmol, 1.00 equiv), (2-methylpropyl)boronic acid (168 mg,1.65 mmol, 1.50 equiv), K₃PO₄-3H₂O (877 mg, 3.00 equiv) and A-Phos-PdCl₂(77.8 mg, 0.10 equiv) in ethylene glycol dimethyl ether (20 mL) and H₂O(2 mL) was stirred under nitrogen at 100° C. overnight. The resultingmixture was cooled to room temperature and concentrated under vacuum.The residue was purified by Prep-HPLC with the following conditions(1#-Pre-HPLC-005 (Waters)): Column, XBridge Shield RP18 OBD Column, 5μm, 19×150 mm; mobile phase, water with 10 mmol NH₄HCO₃ and CH₃CN (18%CH₃CN up to 58% in 10 min, up to 95% in 1 min, down to 18% in 2 min);Detector, UV 254/220 nm to give 50 mg (15%) of tert-butyl2-(((3-iso-butyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamateas a colorless oil. LCMS (method A, ESI): RT=1.27 min, m/z=395.0 [M+H]⁺.

Step 2:N¹-((3-iso-butyl-1H-pyrazol-4-yl)methyl)-N¹-methylethane-1,2-diamine(Compound 28)

A solution of tert-butyl2-(((3-iso-butyl-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate(50 mg, 0.13 mmol, 1.00 equiv) in THF (10 mL) and 12N hydrochloric acid(2 mL) was stirred overnight at 25° C. The resulting mixture wasconcentrated under vacuum. The residue was diluted with 5 mL oftetrahydrofuran and the pH value of the solution was adjusted to 9 with10% sodium carbonate solution. The resulting mixture was concentratedunder vacuum and the residue was dissolved in 5 mL of methanol thenpurified by Prep-HPLC with the following conditions (1#-Pre-HPLC-005(Waters)): Column, XBridge Shield RP18 OBD Column, 5 μm, 19×150 mm;mobile phase, water with 10 mmol NH₄HCO₃ and CH₃CN (18% CH₃CN up to 58%in 10 min, up to 95% in 1 min, down to 18% in 2 min); Detector, UV254/220 nm to yield 6 mg (23%) ofN¹-((3-iso-butyl-1H-pyrazol-4-yl)methyl)-N¹-methylethane-1,2-diamine asa light yellow oil. ¹H-NMR (300 MHz, CD₃OD) 7.49 (s, 1H), 3.44 (s, 2H),2.84-2.80 (m, 2H), 2.56-2.50 (m, 4H), 2.21 (s, 3H), 2.03-1.93 (m, 1H),0.95-0.92 (m, 6H) ppm. LCMS (method AA1 ESI): RT=1.02 min, m/z=211.0[M+H]⁺.

Compound 37N¹-methyl-N¹-((3-(4-methylcyclohexyl)-1H-pyrazol-4-yl)methyl)ethane-1,2-diamine

Step 1: (R/S) tert-butyl2-(methyl((3-(4-methylcyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate

A mixture of (R/S) tert-butylN-[2-([[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl](methyl)amino)ethyl]carbamate(50 mg, 0.11 mmol, 1.00 equiv), potassium carbonate (45 mg, 0.33 mmol,3.02 equiv),4,4,5,5-tetramethyl-2-(4-methylcyclohex-1-en-1-yl)-1,3,2-dioxaborolane(36 mg, 0.16 mmol, 1.51 equiv), Pd(dppf)Cl₂ (8 mg, 0.01 mmol, 0.10equiv) in water (1 mL) and 1,4-dioxane (10 mL) was stirred undernitrogen at 100° C. overnight. The reaction mixture was cooled to roomtemperature and concentrated under vacuum. The residue was purified on asilica gel column eluted with 0-50% of ethyl acetate in petroleum etherto give 30 mg (64%) of (R/S) tert-butyl2-(methyl((3-(4-methylcyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamateas a brown oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.29 (s, 1H), 6.14-6.13 (m,1H), 5.36-5.32 (m, 1H), 4.16-4.07 (m, 2H), 3.70-3.27 (m, 2H), 2.54-2.29(m, 6H), 2.54-2.29 (m, 4H), 2.22 (s, 3H), 2.13-2.07 (m, 3H), 1.86-1.56(m, 4H), 1.47 (s, 9H), 1.46-1.38 (m, 3H) ppm. LCMS (method A, ESI):RT=1.31 min, m/z=433.0 [M+H]⁺.

Step 2: (R/S) tert-butyl2-(methyl((3-(4-methylcyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate

A mixture of (R/S) tert-butyl2-(methyl((3-(4-methylcyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate(200 mg, 0.46 mmol, 1.00 equiv) and 10% palladium on carbon (30 mg)catalyst in methanol (20 mL) was stirred under 20 atm of hydrogen in a50-mL high pressure reactor at 25° C. for 2 days. The catalyst wasremoved by filtration and the filtrate was concentrated under vacuum togive 200 mg of crude (R/S) tert-butyl2-(methyl((3-(4-methylcyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamateas a yellow oil. The crude product was used in the next step withoutfurther purification. LCMS (method C, ESI): RT=0.77 min, m/z=435.0[M+H]⁺.

Step 3:N¹-methyl-N¹-((3-(4-methylcyclohexyl)-1H-pyrazol-4-yl)methyl)ethane-1,2-diamine(Compound 37)

A solution of (R/S) tert-butyl2-(methyl((3-(4-methylcyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate(200 mg, 0.46 mmol, 1.00 equiv) in 4N hydrochloric acid (10 mL) wasstirred at 60° C. for 2 h. The resulting mixture was cooled to roomtemperature and concentrated under vacuum. The crude product waspurified by Prep-HPLC with the following conditions (2#-Waters2767-2(HPLC-08)): Column, XBridge Shield RP 18, 5 μm, 19×150 mm; mobilephase, water with 50 mmol CF₃COOH and CH₃CN (10.0% CH₃CN up to 28.0% in2 min, up to 46.0% in 10 min, up to 100.0% in 1 min, down to 10.0% in 1min); Detector, UV 254 nm to yield 62.3 mg (28%) ofN¹-methyl-N¹-((3-(4-methylcyclohexyl)-1H-pyrazol-4-yl)methyl)ethane-1,2-diaminetrifluoroacetate as a colorless semi-solid. ¹H-NMR (300 MHz, D₂O): δ7.78 (s, 1H), 4.28 (s, 2H), 3.47-3.31 (m, 4H), 2.79-2.60 (s, 4H),2.74-2.70 (m, 1H), 1.90-1.25 (m, 8H), 0.89 (d, J=7.2 Hz, 3H) ppm. LCMS(method V, ESI): RT=1.51 min, 9.12 min, m/z=251.1 [M+H]⁺.

Compound 38N¹-((3-(4,4-dimethylcyclohexyl)-1H-pyrazol-4-yl)methyl)-N¹-methylethane-1,2-diamine

Step 1: (R/S) tert-butyl2-(((3-(4,4-dimethylcyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate

A mixture of (R/S) tert-butylN-[2-([[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl](methyl)amino)ethyl]carbamate(300 mg, 0.65 mmol, 1.00 equiv), Pd(dppf)Cl₂ (52 mg, 0.07 mmol, 0.11equiv),2-(4,4-dimethylcyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(229 mg, 0.97 mmol, 1.50 equiv) and potassium carbonate (268 mg, 1.94mmol, 3.00 equiv) in 1,4-dioxane (20 mL) and water (4 mL) was stirredunder nitrogen at 100° C. overnight. The resulting mixture was cooled toroom temperature and concentrated under vacuum. The residue was purifiedon a silica gel column eluted with 1-41% of ethyl acetate in petroleumether to give 250 mg (87%) of (R/S) tert-butyl2-(((3-(4,4-dimethylcyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamateas a yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.50 (s, 1H), 6.14-6.13 (m,1H), 5.36-5.32 (m, 1H), 4.18-4.07 (m, 2H), 3.74-3.67 (m, 1H), 3.41-3.25(m, 4H), 2.51-2.50 (m, 3H), 2.20-2.02 (m, 6H), 1.73-1.71 (m, 3H),1.70-1.66 (m, 6H), 1.47 (s, 9H), 1.28-1.26 (m, 4H) ppm. LCMS (method D,ESI): RT=1.33 min, m/z=447.0 [M+H]⁺.

Step 2: (R/S) tert-butyl2-(((3-(4,4-dimethylcyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate

A mixture of (R/S) tert-butyl2-(((3-(4,4-dimethylcyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethylcarbamate(250 mg, 0.56 mmol, 1.00 equiv) and 10% palladium on carbon (30 mg)catalyst in methanol (20 mL) was stirred under 20 atm. of hydrogen in a50-mL high pressure reactor at 25° C. for 2 days. The catalyst wasremoved by filtration. The filtrate was concentrated under vacuum togive 250 mg of crude (R/S) tert-butyl2-(((3-(4,4-dimethylcyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamateas a yellow oil. The crude product was used in the next step withoutfurther purification. LCMS (method C, ESI): RT=0.80 min, m/z=449.0[M+H]⁺.

Step 3:N¹-((3-(4,4-dimethylcyclohexyl)-1H-pyrazol-4-yl)methyl)-N¹-methylethane-1,2-diamine(Compound 38)

A solution of (R/S) tert-butyl2-(((3-(4,4-dimethylcyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)carbamate(250 mg, 0.56 mmol, 1.00 equiv) in 4N hydrochloric acid (10 mL) wasstirred at 60° C. for 2 h. The reaction mixture was cooled to roomtemperature and concentrated under vacuum. The crude product waspurified by Prep-HPLC with the following conditions (2#-Waters 2767-2(HPLC-08)): Column, XBridge Shield RP 18, 5 μm, 19×150 mm; mobile phase,water with 50 mmol CF₃COOH and CH₃CN (10.0% CH₃CN up to 28.0% in 2 min,up to 46.0% in 10 min, up to 100.0% in 1 min, down to 10.0% in 1 min);Detector, UV 254 nm to yield 171.2 mg (62%) ofN¹-((3-(4,4-dimethylcyclohexyl)-1H-pyrazol-4-yl)methyl)-N¹-methylethane-1,2-diaminetrifluoroacetate as a light yellow oil. ¹H-NMR (300 MHz, D₂O): δ 7.75(s, 1H), 4.30 (s, 2H), 3.47-3.35 (m, 4H), 2.77 (s, 3H), 2.68-2.58 (m,1H), 1.71-1.53 (m, 4H), 1.49-1.37 (m, 2H), 1.31-1.17 (m, 2H), 1.89 (s,3H), 1.87 (s, 3H) ppm. LCMS (method M, ESI): RT=1.15, m/z=265.1 [M+H]⁺.

Compound 39N¹-((3-(1-isobutylpiperidin-4-yl)-1H-pyrazol-4-yl)methyl)-N¹,N²-dimethylethane-1,2-diamine

Step 1: (R/S) tert-butyl2-(((3-(1-isobutylpiperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)(methyl)carbamate

To a solution of (R/S) tert-butylN-methyl-N-[2-[methyl([[1-(oxan-2-yl)-3-(piperidin-4-yl)-1H-pyrazol-4-yl]methyl])amino]ethyl]carbamate(250 mg, 0.57 mmol, 1.00 equiv) and 2-methylpropanal (62 mg, 0.86 mmol,1.50 equiv) in 1,2-dichloroethane (15 mL) was added NaBH(OAc)₃ (364 mg,3.00 equiv). The resulting solution was stirred at room temperatureovernight and then concentrated under vacuum to give 160 mg of crude(R/S) tert-butyl2-(((3-(1-isobutylpiperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl(methyl)carbamateas a light yellow oil. LCMS (method A, ESI): RT=1.52 min, m/z=492.2[M+H]⁺.

Step 2:N¹-((3-(1-isobutylpiperidin-4-yl)-1H-pyrazol-4-yl)methyl)-N¹,N²-dimethylethane-1,2-diamine(Compound 39)

A solution of tert-butyl2-(((3-(1-isobutylpiperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl)(methyl)carbamate(130 mg, 0.26 mmol, 1.00 equiv) in ethanol (2 mL), 1,4-dioxane (4 mL)and 3N hydrochloric acid (2 mL) was stirred at room temperatureovernight. The reaction mixture was concentrated under vacuum and theresidue was purified by Pre-HPLC with the following conditions(1#-Pre-HPLC-005(Waters)): Column, SunFire Prep C18 OBD Column, 5 μm,19×150 mm; mobile phase, phase A: water with 0.05% TFA; phase B: MeCN(5% CH₃CN up to 17% in 10 min, down to 0% in 0 min); Detector, UV254/220 nm to give 39.5 mg (28%) ofN¹-((3-(1-isobutylpiperidin-4-yl)-1H-pyrazol-4-yl)methyl)-N¹,N²-dimethylethane-1,2-diaminetrifluoroacetate as a colorless solid. ¹H-NMR (300 MHz, D₂O): δ 7.81 (s,1H), 4.32 (s, 2H), 3.71-3.35 (m, 7H), 3.15-2.89 (m, 4H), 2.82-2.68 (m,6H), 2.22-1.92 (m, 5H), 0.93 (d, J=6.8 Hz, 6H) ppm. LCMS (method U,ESI): m/z=308.2 [M+H]⁺.

Compound 403-methyl-1-(4-(4-((methyl(2-(methylamino)ethyl)amino)methyl)-1H-pyrazol-3-yl)piperidin-1-yl)butan-1-one

Step 1: (R/S) benzyl4-(4-(((2-(tert-butoxycarbonyl(methyl)amino)ethyl)(methyl)amino)methyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)-5,6-dihydropyridine-1(2H)-carboxylate

A mixture of (R/S) tert-butylN-[2-([[4-iodo-1-(oxan-2-yl)-1H-pyrrol-3-yl]methyl](methyl)amino)ethyl]-N-methylcarbamate(3.15 g, 6.60 mmol, 1.00 equiv), benzyl4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine-1-carboxylate(2.5 g, 7.28 mmol, 1.10 equiv), Pd(dppf)Cl₂ (1.39 g, 1.90 mmol, 0.29equiv) and potassium carbonate (2.72 g, 19.68 mmol, 2.98 equiv) in1,4-dioxane (30 mL) and water (3 mL) was stirred under nitrogen at 100°C. overnight. The reaction was cooled to room temperature and thenquenched by the addition of 30 mL of water. The resulting mixture wasextracted with 3×250 mL of ethyl acetate. The combined organic layerswas dried over anhydrous sodium sulfate and concentrated under vacuum.The residue was purified on a silica gel column eluted with 0-15% ofethyl acetate in petroleum ether to give 2.1 g (56%) of (R/S) benzyl4-(4-(((2-(tert-butoxycarbonyl(methyl)amino)ethyl)(methyl)amino)methyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)-5,6-dihydropyridine-1(2H)-carboxylateas a yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.58-7.49 (m, 1H), 7.49-7.35(m, 4H), 7.35-7.30 (m, 1H), 5.33-5.30 (m, 1H), 5.20 (s, 2H), 4.25-4.00(m, 3H), 3.70-3.69 (m, 3H), 3.39-3.31 (m, 3H), 2.84 (m, 3H), 2.66 (m,2H), 2.50 (m, 2H), 2.25 (m, 2H), 2.08-2.07 (m, 3H), 1.73-1.62 (m, 4H),1.46 (s, 9H), 1.31-1.27 (m, 1H) ppm. LCMS (method A, ESI): RT=0.74 min,m/z=568.0 [M+H]⁺.

Step 2: (R/S) tert-butylmethyl(2-(methyl((3-(piperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate

A mixture of benzyl4-(4-(((2-(tert-butoxycarbonyl(methyl)amino)ethyl)(methyl)amino)methyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)-5,6-dihydropyridine-1(2H)-carboxylate(2 g, 3.52 mmol, 1.00 equiv) and 10% palladium on carbon (2 g) catalystin methanol (100 mL) was stirred under 1 atmosphere of hydrogen at roomtemperature for 6 h. The catalyst was removed by filtration and thefiltrate was concentrated under vacuum to yield 1.1 g (72%) of (R/S)tert-butylmethyl(2-(methyl((3-(piperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamateas a brown oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.46 (s, 1H), 5.33-5.32 (m,1H), 4.24-4.06 (m, 1H), 3.75-3.66 (m, 1H), 3.51 (s, 1H), 3.41-3.15 (m,6H), 2.95-2.70 (m, 6H), 2.62-2.40 (m, 2H), 2.22 (s, 3H), 1.55-1.41 (m,10H), 1.35-1.21 (m, 1H) ppm. LCMS (method A, ESI): RT=1.49 min,m/z=436.2 [M+H]⁺.

Step 3: (R/S) tert-butylmethyl(2-(methyl((3-(1-(3-methylbutanoyl)piperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate

To a solution of (R/S) tert-butylmethyl(2-(methyl((3-(piperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate(200 mg, 0.46 mmol, 1.00 equiv) and triethylamine (1.14 g, 11.26 mmol,24.52 equiv) in dichloromethane (15 mL) was added 3-methylbutanoylchloride (67 mg, 0.56 mmol, 1.21 equiv). The resulting solution wasstirred at room temperature for 2 h. The reaction was then quenched bythe addition of 2 mL of water. The organic layer was dried overanhydrous sodium sulfate and concentrated under vacuum to give 250 mg ofcrude (R/S) tert-butylmethyl(2-(methyl((3-(1-(3-methylbutanoyl)piperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamateas a yellow solid. LCMS (method D, ESI): RT=1.22 min, m/z=520.0 [M+H]⁺.

Step 4:3-methyl-1-(4-(4-((methyl(2-(methylamino)ethyl)amino)methyl)-1H-pyrazol-3-yl)piperidin-1-yl)butan-1-one(Compound 40)

A solution of tert-butylmethyl(2-(methyl((3-(1-(3-methylbutanoyl)piperidin-4-yl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate(110 mg, 0.21 mmol, 1.00 equiv) in ethanol (2 mL), 1,4-dioxane (4 mL)and 12N hydrochloric acid (2 mL) was stirred at room temperatureovernight. The resulting mixture was concentrated under vacuum and theresidue was purified by Pre-HPLC with the following conditions(1#-Pre-HPLC-005 (Waters)): Column, XBridge Shield RP18 OBD Column, 5μm, 19×150 mm; mobile phase, water with 10 mmol NH₄HCO₃ and CH₃CN (18%CH₃CN up to 58% in 10 min, up to 95% in 1 min, down to 18% in 2 min);Detector, UV 254/220 nm to give 17.7 mg (25%) of3-methyl-1-(4-(4-((methyl(2-(methylamino)ethyl)amino)methyl)-1H-pyrazol-3-yl)piperidin-1-yl)butan-1-oneas a colorless solid. ¹H-NMR (300 MHz, D₂O): δ 7.53 (s, 1H), 4.50-4.40(m, 1H), 4.10-4.00 (m, 1H), 3.44 (s, 2H), 3.25-3.10 (m, 1H), 3.09-2.95(m, 1H), 2.80-2.65 (m, 3H), 2.53-2.43 (m, 2H), 2.40-2.20 (m, 5H), 2.13(s, 3H), 2.00-1.75 (m, 3H), 1.72-1.43 (m, 2H), 0.88 (d, J=6.8 Hz, 6H)ppm. LCMS (method R, ESI): RT=1.26 min, m/z=336.2 [M+H]⁺.

Compound 43N¹-methyl-N¹-((3-(4-morpholinocyclohexyl)-1H-pyrazol-4-yl)methyl)ethane-1,2-diamine

Step 1: (R/S) tert-butyl2-(methyl((3-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate

A mixture of (R/S) tert-butylN-[2-([[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl](methyl)amino)ethyl]carbamate(400 mg, 0.86 mmol, 1.00 equiv), Pd(dppf)Cl₂ (66 mg, 0.09 mmol, 0.10equiv), potassium carbonate (356 mg, 2.58 mmol, 2.99 equiv) and4-[4-(tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl]morpholine(379 mg, 1.29 mmol, 1.50 equiv) in 1,4-dioxane (20 mL) and water (2 mL)was stirred under nitrogen at 100° C. overnight. The resulting mixturewas cooled to room temperature then concentrated under vacuum. Theresidue was purified on a silica gel column eluted with 0-3% of methanolin dichloromethane to give 320 mg (74%) of (R/S) tert-butyl2-(methyl((3-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamateas a brown oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.52 (s, 1H), 6.13-6.12 (m,1H), 5.35-5.31 (m, 1H), 4.18-4.11 (m, 2H), 3.80-3.78 (m, 5H), 3.45-3.43(m, 2H), 3.26-3.25 (m, 2H), 2.69-2.63 (m, 5H), 2.54-2.48 (m, 4H),2.25-2.20 (m, 4H), 2.13-2.02 (m, 4H), 1.67-1.61 (m, 4H), 1.47 (s, 9H)ppm. LCMS (method A, ESI): RT=1.00 min, m/z=504.0 [M+H]⁺.

Step 2: (R/S) tert-butyl2-(methyl((3-(4-morpholinocyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate

A mixture of (R/S) tert-butyl2-(methyl((3-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate(300 mg, 0.60 mmol, 1.00 equiv) and 10% palladium on carbon (20 mg)catalyst in acetic acid (15 mL) was stirred under 20 atm of hydrogen ina 50-mL high pressure reactor at 25° C. for 3 days. The catalyst wasremoved by filtration and the filtrate was concentrated under vacuum togive 300 mg of crude (R/S) tert-butyl2-(methyl((3-(4-morpholinocyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamateas a yellow oil. The crude product was used in the next step withoutfurther purification. LCMS (method A, ESI): RT=1.01 min, m/z=506.0[M+H]⁺.

Step 3:N¹-methyl-N¹-((3-(4-morpholinocyclohexyl)-1H-pyrazol-4-yl)methyl)ethane-1,2-diamine(Compound 43)

A solution of (R/S) tert-butyl2-(methyl((3-(4-morpholinocyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methyl)amino)ethyl)carbamate(300 mg, 0.59 mmol, 1.00 equiv) in 4N hydrochloric acid (15 mL) wasstirred at 60° C. for 2 h. The resulting mixture was cooled to roomtemperature then concentrated under vacuum. The crude product waspurified by Prep-HPLC with the following conditions (Waters2767-2(HPLC-08)): Column, XBridge Shield RP 18, 5 μm, 19×150 mm; mobilephase, water with 50 mmol CF₃COOH and CH₃CN (10.0% CH₃CN up to 28.0% in2 min, up to 46.0% in 10 min, up to 100.0% in 1 min, down to 10.0% in 1min); Detector, UV 254 nm to afford 36.5 mg (11%) ofN¹-methyl-N¹-((3-(4-morpholinocyclohexyl)-1H-pyrazol-4-yl)methyl)ethane-1,2-diaminetrifluoroacetate as a white solid. ¹H-NMR (300 MHz, CD₃OD): δ 7.47 (s,1H), 3.78-3.71 (m, 4H), 3.47 (s, 2H), 2.88-2.80 (m, 2H), 2.80-2.70 (m,1H), 2.70-2.62 (m, 4H), 2.57-2.50 (m, 2H), 2.45-2.27 (m, 1H), 2.23 (s,3H), 2.16-1.93 (m, 4H), 1.75-1.57 (m, 2H), 1.50-1.34 (m, 2H) ppm. LCMS(method M, ESI): m/z=322.2 [M+H]⁺.

Compound 44N¹-methyl-N¹-((4-(4-morpholinocyclohexyl)-1H-pyrazol-3-yl)methyl)ethane-1,2-diamine

Step 1: 4-morpholinocyclohex-1-enyl trifluoromethanesulfonate

To a stirred solution of 4-(morpholin-4-yl)cyclohexan-1-one (920 mg,5.02 mmol, 1.00 equiv) in anhydrous tetrahydrofuran (20 mL) maintainedunder nitrogen at −78° C. was added dropwise a 1M solution of LiHMDS (6mL) in tetrahydrofuran. After stirring for 1 h at −78° C., a solution of1,1,1-trifluoro-N-phenyl-N-(trifluoromethane)-sulfonylmethane-sulfonamide(1.97 g, 5.51 mmol, 1.10 equiv) in tetrahydrofuran (6 mL) was added. Thereaction was warmed to room temperature and stirred for 12 h. Theresulting solution was concentrated under vacuum and the residue waspurified on a silica gel column eluted with 50-100% of ethyl acetate inpetroleum ether to give 420 mg (27%) of 4-morpholinocyclohex-1-enyltrifluoromethanesulfonate as a yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ5.80-5.70 (m, 1H), 3.90-3.75 (m, 4H), 2.75-2.00 (m, 10H), 1.70-1.50 (m,1H) ppm.

Step 2:4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)morpholine

A mixture of 4-morpholinocyclohex-1-enyl trifluoromethanesulfonate (4 g,12.69 mmol, 1.00 equiv),4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(3.87 g, 15.24 mmol, 1.20 equiv), potassium acetate (3.73 g, 38.01 mmol,3.00 equiv) and Pd(dppf)Cl₂ (930 mg, 1.27 mmol, 0.10 equiv) in1,4-dioxane (100 mL) was refluxed under nitrogen for 12 h. The reactionmixture was cooled to room temperature, filtered and then concentratedunder vacuum. The residue was purified on a silica gel column elutedwith 50-100% of ethyl acetate in petroleum ether to give 3.2 g (86%) of4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)morpholineas a yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 6.60-6.55 (m, 1H), 3.80-3.66(m, 4H), 2.70-2.25 (m, 8H), 2.20-1.90 (m, 4H), 1.25 (s, 12H) ppm.

Step 3: (R/S)4-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-3-carbaldehyde

A mixture of4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)morpholine(293 mg, 1.00 mmol, 1.00 equiv), (R/S)4-iodo-1-(oxan-2-yl)-1H-pyrazole-3-carbaldehyde (306 mg, 1.00 mmol, 1.00equiv), K₃PO₄ (640 mg, 3.02 mmol, 3.02 equiv) and Pd(dppf)Cl₂ (65.1 mg,0.10 mmol, 0.10 equiv) in ethylene glycol dimethyl ether (5 mL) wasstirred under nitrogen at 85° C. for 12 h. The reaction was cooled toroom temperature and concentrated under vacuum. The residue was purifiedon a silica gel column eluted with 50-100% of ethyl acetate in petroleumether to give 280 mg (81%) of (R/S)4-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-3-carbaldehydeas a brown oil. LCMS (method C, ESI): RT=0.70 min, m/z=346.2 [M+H]⁺.

Step 4: (R/S) tert-butyl2-(methyl((4-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methyl)amino)ethyl)carbamate

To a solution of (R/S)4-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-3-carbaldehyde(500 mg, 1.45 mmol, 1.00 equiv) and tert-butylN-[2-(methylamino)ethyl]carbamate (378 mg, 2.17 mmol, 1.50 equiv) in1,2-dichloroethane (20 mL) was added NaBH(OAc)₃ (612 mg, 2.89 mmol, 1.99equiv). The reaction mixture was stirred at room temperature for 12 hand then quenched with saturated NaHCO₃ solution (10 mL). The organiclayer was dried over anhydrous sodium sulfate and concentrated undervacuum. The residue was purified on a silica gel column eluted with20-100% of ethyl acetate in petroleum ether to give 300 mg (41%) of(R/S) tert-butyl2-(methyl((4-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methyl)amino)ethyl)carbamateas a brown oil. LCMS (method A, ESI): RT=0.66 min, m/z=504.4 [M+H]⁺.

Step 5: (R/S) tert-butyl2-(methyl((4-(4-morpholinocyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methyl)amino)ethyl)carbamate

A mixture of (R/S) tert-butyl2-(methyl((4-(4-morpholinocyclohex-1-enyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methyl)amino)ethyl)carbamate(252 mg, 0.50 mmol, 1.00 equiv) and 10% palladium on carbon catalyst (25mg) in acetic acid (10 mL) was stirred in a 30-mL pressure reactor under20 atm. of hydrogen at 25° C. for 12 h. The catalyst was removed byfiltration and the filtrate was concentrated to give 250 mg (99%) of(R/S) tert-butyl2-(methyl((4-(4-morpholinocyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methyl)amino)ethyl)carbamateas a yellow oil. LCMS (method C, ESI): RT=0.66 min, m/z=506.4 [M+H]⁺.

Step 6:N¹-methyl-N¹-((4-(4-morpholinocyclohexyl)-1H-pyrazol-3-yl)methyl)ethane-1,2-diamine(Compound 44)

A mixture of (R/S) tert-butyl2-(methyl((4-(4-morpholinocyclohexyl)-1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-3-yl)methyl)amino)ethyl)carbamate(253 mg, 0.50 mmol, 1.00 equiv) in a saturated solution of hydrogenchloride in 1,4-dioxane (20 mL) was stirred at 25° C. for 24 h. Theresulting mixture was concentrated under vacuum and the crude product(150 mg) was purified by Prep-HPLC with the following conditions(Prep-HPLC-005): Column, XBridge Prep C18 OBD Column, 5 m, 19×150 mm;mobile phase, water with 10 mmol NH₄HCO₃ and MeCN (hold 4% MeCN in 5min, up to 5% in 10 min); Detector, UV 254/220 nm to give 30 mg (19%) ofN-methyl-N-((4-(4-morpholinocyclohexyl)-1H-pyrazol-3-yl)methyl)ethane-1,2-diamineas a colorless oil. ¹H-NMR (300 MHz, CD₃OD): δ 7.40 (s, 1H), 3.75-3.65(m, 4H), 3.58 (s, 2H), 2.80-2.72 (m, 2H), 2.69-2.27 (m, 8H), 2.19 (s,3H), 2.12-1.93 (m, 4H), 1.55-1.28 (m, 4H) ppm. LCMS (method W):m/z=322.2 [M+H]⁺.

Compound 106 Methyl[2-(methylamino)ethyl]([3-[(5R,8R)-1,1-dimethyl-2-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amine

Step 1: 1,4,10-trioxadispiro[4.2.48.25]tetradecan-9-one

Into a 500-mL 3-necked round-bottom flask was placed THF (150 mL), LDA(1.3 equiv, prepared from 36 mL of n-BuLi (2.5 M in hexane) reacted with13.8 mL of diisopropylamine for 30 min at −50° C.). Then ethyl1,4-dioxaspiro[4.5]decane-8-carboxylate (15 g, 70.01 mmol, 1.0 equiv)was added and stirred for 30 min at −70° C., followed by oxirane (0.22g/mL in THF, 28 mL) at −78° C. The resulting solution was stirred for 2h at −70° C. The reaction was quenched by 100 mL of NH₄Cl (sat. aq.),then treated with 100 mL of EtOAc. The organic phase was separated andthen washed with 150 mL of brine. The organic phase was dried andconcentrated under vacuum. The residue was purified by flashchromatography on silica gel using ethyl acetate/petroleum ether (1:2)as eluent to give 4.5 g (30%) of1,4,10-trioxadispiro[4.2.4⁸.2⁵]tetradecan-9-one as a yellow solid.¹H-NMR (300 MHz, CDCl₃): δ 4.28 (t, J=6.6 Hz, 2H), 4.10-3.85 (m, 4H),2.17 (t, J=6.6 Hz, 2H), 2.15-1.85 (m, 4H), 1.80-1.50 (m, 4H).

Step 2: 2-[8-(2-hydroxyethyl)-1,4-dioxaspiro[4.5]decan-8-yl]propan-2-ol

To a stirred solution of 1,4,10-trioxadispiro[4.2.4⁸.2⁵]tetradecan-9-one(3.18 g, 14.98 mmol, 1.0 equiv) in THF (100 mL) at 0° C. was addeddropwise a solution of MeMgBr (1M in ether, 75 mL, 5.0 equiv). Theresulting solution was allowed to warm to room temperature and stirredfor 12 h at room temperature. The reaction was quenched with 40 mL ofNH₄Cl (sat. aq.), then treated with 300 mL of EtOAc. The organic phasewas separated and then washed with 100 mL brine then dried withanhydrous Na₂SO₄. Concentration under reduced pressure afforded 4.9 g(crude) of2-[8-(2-hydroxyethyl)-1,4-dioxaspiro[4.5]decan-8-yl]propan-2-ol asyellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 4.00-3.90 (m, 4H), 3.80-3.40 (m,4H), 1.95-1.50 (m, 10H), 1.40-1.10 (m, 6H).

Step 3: 9,9-dimethyl-1,4,10-trioxadispiro[4.2.48.25]tetradecane

Into a 250-mL round-bottom flask was placed2-[8-(2-hydroxyethyl)-1,4-dioxaspiro[4.5]decan-8-yl]propan-2-ol (4.91 g,20.10 mmol, 1.00 equiv), dichloromethane (60 mL),4-dimethylaminopyridine (300 mg, 2.46 mmol, 0.12 equiv) andtriethylamine (20 mL). Tosylchloride (5.34 g, 28.01 mmol, 1.39 equiv)was added and the resulting solution was stirred for 12 h at roomtemperature then concentrated to dryness under reduced pressure. Theresidue was purified by flash chromatography on silica gel using ethylacetate/petroleum ether (1:2) as eluent to give 3.5 g (77%) of9,9-dimethyl-1,4,10-trioxadispiro[4.2.4⁸.2⁵]tetradecane as a yellow oil.¹H-NMR (300 MHz, CDCl₃): δ 3.84 (s, 4H), 3.66 (t, J=7.5 Hz, 2H), 2.51(t, J=7.5 Hz, 2H), 1.70-1.25 (m, 8H), 0.99 (s, 6H).

Step 4: 1,1-dimethyl-2-oxaspiro[4.5]decan-8-one

Into a 100-mL round-bottom flask purged and maintained with anatmosphere of nitrogen was placed9,9-dimethyl-1,4,10-trioxadispiro[4.2.4⁸.2⁵]tetradecane (1.765 g, 7.80mmol, 1.00 equiv), tetrahydrofuran (16 mL) and hydrochloric acid (12N,16 mL). The resulting solution was stirred for 6 h at room temperaturethen extracted with 3×50 mL of ethyl acetate. The combined organiclayers were dried over anhydrous sodium sulfate and concentrated undervacuum to afford 1.259 g (89%) of1,1-dimethyl-2-oxaspiro[4.5]decan-8-one as a light yellow solid.

Step 5: 1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yltrifluoromethanesulfonate

To a solution of 1,1-dimethyl-2-oxaspiro[4.5]decan-8-one (1.695 g, 9.30mmol, 1.00 equiv) in THF (50 mL) at −70° C. under dry nitrogen was addeddropwise a solution of LiHMDS (1M in THF, 14 mL). The reaction mixturewas stirred for 1 h at −70° C. then treated with1,1,1-trifluoro-N-phenyl-N-(trifluoromethane) sulfonylmethanesulfonamide(3.490 g, 9.77 mmol, 1.05 equiv) and stirred at −70° C. for another 0.5h. The resulting solution was allowed to warm to room temperature andstirred for another 12 hours then concentrated under vacuum. The residuewas purified by flash chromatography on silica using ethylacetate/petroleum ether (2:1) as eluent to afford 2.458 g (84%) of1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate as acolorless oil. ¹H-NMR (300 MHz, CDCl₃): δ 5.80-5.70 (m, 1H), 3.95-3.80(m, 2H), 2.50-2.35 (m, 2H), 2.30-2.15 (m, 1H), 2.10-1.50 (m, 5H).

Step 6:2-[1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen was placed1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate(1.472 g, 4.68 mmol, 1.00 equiv),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1.786 g),1,4-dioxane (20 mL), potassium acetate (1.378 g, 14.04 mmol, 3.00 equiv)and PdCl₂(dppf) (343 mg). The resulting solution was stirred for 12 h at100° C. then quenched by the addition of 10 mL of water/ice. Theresulting solution was extracted with 200 mL of ethyl acetate and thecombined organic layers washed with 50 mL of brine, dried over anhydroussodium sulfate and concentrated under vacuum. The residue was purifiedby flash chromatography on silica gel using ethyl acetate/petroleumether (2:1) as eluent to afford 715 mg (52%) of2-[1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolaneas a white solid.

Step 7: (R/S) tert-butylN-(2-[[(3-[1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen was placed2-[1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(1.164 g, 3.98 mmol, 1.00 equiv), tert-butylN-[2-([[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl](methyl)amino)ethyl]-N-methylcarbamate (2.096 g, 4.38 mmol, 1.10 equiv),potassium carbonate (1.650 g, 11.94 mmol, 3.00 equiv), 1,4-dioxane (20mL), water (2 mL) and PdCl₂(dppf) (292 mg). The resulting solution wasstirred for 16 h at 100° C. then quenched by the addition of 10 mL ofwater/ice. The resulting solution was extracted with 100 mL of ethylacetate and the organic layer separated and washed with 50 mL of brine,dried over anhydrous sodium sulfate and then concentrated under vacuum.The residue was purified by flash chromatography on silica gel columnusing ethyl acetate/petroleum ether (1:1) as eluent to afford 1.081 g(53%) of (R/S) tert-butylN-(2-[[(3-[1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas a yellow oil.

Step 8 (R/S) tert-butylN-(2-[[(3-[1,1-dimethyl-2-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen was placed (R/S) tert-butylN-(2-[[(3-[1,1-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(1.081 g, 2.09 mmol, 1.00 equiv), tetrahydrofuran (25 mL), and 10%Pd(OH)₂/C (400 mg, 2.85 mmol, 1.36 equiv). The resulting solution wasstirred for 5 h at room temperature under 3 atmospheres of hydrogen. Theresulting mixture was filtered and the filtrate concentrated undervacuum to afford 860 mg (79%) of (R/S) tert-butylN-(2-[[(3-[1,1-dimethyl-2-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas a light yellow oil.

Step 9:Methyl[2-(methylamino)ethyl]([3-[(5R,8S)-1,1-dimethyl-2-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amine(Compound 106)

A solution of (R/S) tert-butylN-(2-[[(3-[1,1-dimethyl-2-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(400 mg, 1.20 mmol, 1.00 equiv) in methanol (6 mL) was treated withhydrochloric acid (12N, 2 mL) and stirred for 1 h at room temperaturethen for an additional 2 h at 50° C. The reaction was then quenched bythe addition of 20 mL of sodium bicarbonate (sat. aq.) and the resultingmixture concentrated under vacuum to remove the majority of themethanol. The resulting solution was extracted with 2×30 mL ofdichloromethane and the organic layers combined and dried over anhydroussodium sulfate and concentrated under vacuum. The crude product (200 mg)was purified by Chiral prep-HPLC. Column: IC4.6×100 nm, Size: 0.46×10cm, 5 m; Mobile phase: Hex (0.2% IPA): IPA=85:15; Flow: 1.0 ml/min;Detector: UV-220 nm; Instrument: LC-05; Temperature: 25° C. Thisresulted in 32.6 mg ofmethyl[2-(methylamino)ethyl]([3-[(5R,8R)-1,1-dimethyl-2-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amineas a colorless solid. ¹H-NMR (300 MHz, D₂O): δ 7.50 (s, 1H), 3.76 (t,J=7.5 Hz, 2H), 3.42 (s, 2H), 2.80-2.45 (m, 5H), 2.31 (s, 3H), 2.12 (s,3H), 2.01 (t, J=7.5 Hz, 2H), 1.80-1.25 (m, 8H), 1.05 (s, 6H) ppm. LCMS(method A11, ESI): RT=1.44 min, m/z=335.0 [M+H]⁺.

Compound 133 & 134 Methyl[2-(methylamino)ethyl]([3-[(5s,8s)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amineandmethyl[2-(methylamino)ethyl]([3-[(5r,8r)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amine

Step 1: 3-(benzyloxy)-2,2-dimethylpropan-1-ol

Into a 250-mL round-bottom flask, was placed2,2-dimethylpropane-1,3-diol (10.4 g, 99.86 mmol) andN,N-dimethylformamide (100 mL). This was followed by the addition of 60%sodium hydride (4 g, 100.00 mmol), in portions at 0° C. To this wasadded (bromomethyl)benzene (13.68 g, 79.98 mmol) at 0° C. The resultingsolution was stirred for 12 h at room temperature and then diluted with200 mL of NH₄Cl (sat. aq). The resulting solution was extracted with2×200 mL of ethyl acetate and the organic layers were combined and driedover anhydrous sodium sulfate and concentrated under vacuum. The residuewas purified by silica gel column with ethyl acetate/petroleum ether(1:10) to obtain 12 g (62%) of 3-(benzyloxy)-2,2-dimethylpropan-1-ol aslight yellow oil. ¹H NMR (300 MHz, DMSO-d₆): δ 7.43-7.24 (m, 5H),4.51-4.41 (m, 3H), 3.25-3.15 (m, 4H), 0.84 (s, 6H) ppm.

Step 2: [(3-iodo-2,2-dimethylpropoxy)methyl]benzene

Into a 250-mL round-bottom flask, was placed3-(benzyloxy)-2,2-dimethylpropan-1-ol (4 g, 20.59 mmol), imidazole (2.80g, 41.18 mmol), triphenylphosphine (8.1 g, 30.88 mmol), andtetrahydrofuran (100 mL). This was followed by the addition of iodine(7.85 g, 30.93 mmol) in portions at 0° C. The resulting solution wasstirred for 12 h at room temperature and then for an additional 4 hoursat 80° C. and then concentrated under vacuum. The residue was purifiedby silica gel column with petroleum ether to obtain 6 g (96%) of[(3-iodo-2,2-dimethylpropoxy)methyl]benzene as colorless oil. ¹H NMR(300 MHz, DMSO-d₆): δ 7.42-7.20 (m, 5H), 4.49 (s, 2H), 3.30 (s, 2H),3.24 (s, 2H), 1.00 (s, 6H) ppm.

Step 3:8-[3-(benzyloxy)-2,2-dimethylpropyl]-1,4-dioxaspiro[4.5]decan-8-ol

Into a 250-mL 3-necked round-bottom flask that was maintained with anatmosphere of nitrogen, was placed tetrahydrofuran (30 mL). This wasfollowed by the addition of t-BuLi (1.3M in pentane, 40 mL) dropwisewith stirring at −78° C. To this was added a solution of[(3-iodo-2,2-dimethylpropoxy)methyl]benzene (6.08 g, 20.00 mmol) intetrahydrofuran (30 mL) dropwise with stirring at −78° C. and theresulting mixture was stirred at −78° C. for 1 h. To the mixture wasadded a solution of 1,4-dioxaspiro[4.5]decan-8-one (4.69 g, 30.00 mmol,1.50 equiv) in tetrahydrofuran (30 mL) dropwise with stirring at −78° C.The resulting solution was stirred for 1 h at −78° C., and then waswarmed to room temperature slowly. The reaction mixture was diluted with120 mL of NH₄Cl (sat. aq). The organic layer was collected and theaqueous layer was extracted with 2×100 mL of ethyl acetate and theorganic layers were combined and dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was applied onto a C18 gel columnwith H₂O/CH₃CN (3:7) to obtain 3.5 g (52%) of8-[3-(benzyloxy)-2,2-dimethylpropyl]-1,4-dioxaspiro[4.5]decan-8-ol aslight yellow oil. ¹H NMR (400 MHz, DMSO-d₆): δ 7.42-7.20 (m, 5H), 4.46(s, 2H), 3.92 (s, 1H), 3.82 (s, 4H), 3.21 (s, 2H), 1.80-1.67 (m, 2H),1.66-1.55 (m, 2H), 1.55-1.45 (m, 2H), 1.5-1.35 (m, 4H), 1.00 (s, 6H)ppm.

Step 4: 8-(3-hydroxy-2,2-dimethylpropyl)-1,4-dioxaspiro[4.5]decan-8-ol

Into a 100-mL round-bottom flask, was placed8-[3-(benzyloxy)-2,2-dimethylpropyl]-1,4-dioxaspiro[4.5]decan-8-ol (3.35g, 10.02 mmol), tetrahydrofuran (40 mL), and 10% palladium/carbon (0.34g). This was followed by the addition of formic acid (3.5 mL) dropwisewith stirring. Hydrogen (3 atm) was then applied to the reaction mixtureand the resulting solution stirred for 4 h at room temperature. Thesolids were removed by filtration and the solution was concentratedunder vacuum. The residue was purified by silica gel column with ethylacetate/petroleum ether (1:1) to obtain 1.5 g (61%) of8-(3-hydroxy-2,2-dimethylpropyl)-1,4-dioxaspiro[4.5]decan-8-ol as awhite solid. ¹H NMR (300 MHz, DMSO-d₆): δ 4.85 (t, J=5.4 Hz, 1H), 4.48(s, 1H), 3.82 (s, 4H), 3.17 (d, J=5.4 Hz, 2H), 1.83-1.58 (m, 4H),1.58-1.35 (m, 6H), 0.90 (s, 6H) ppm.

Step 5: 11,11-dimethyl-1,4,9-trioxadispiro[4.2.4̂[8].2̂[5]]tetradecane

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed8-(3-hydroxy-2,2-dimethylpropyl)-1,4-dioxaspiro[4.5]decan-8-ol (4 g,16.37 mmol), tributylphosphane (6.62 g, 32.72 mmol), and tetrahydrofuran(60 mL). A solution of TMAD (5.64 g, 32.75 mmol) in tetrahydrofuran (80mL) was added dropwise with stirring at −40° C. The reaction mixture wasstirred for 1 h at −40° C. and then an additional 12 h at roomtemperature. The resulting mixture was concentrated under vacuum. Theresidue was purified by silica gel column with ethyl acetate/petroleumether (1:4) to obtain 3.2 g (86%) of11,11-dimethyl-1,4,9-trioxadispiro[4.2.4̂[8].2̂[5]]tetradecane ascolorless oil. ¹H NMR (400 MHz, DMSO-d₆): δ3.83 (s, 4H), 3.38 (s, 2H),1.78-1.63 (m, 4H), 1.63-1.42 (m, 6H), 1.03 (s, 6H) ppm.

Step 6: 3,3-dimethyl-1-oxaspiro[4.5]decan-8-one

Into a 100-mL round-bottom flask, was placed11,11-dimethyl-1,4,9-trioxadispiro[4.2.4̂[8].2̂[5]]tetradecane (2.0 g,8.84 mmol, 1.00 equiv), tetrahydrofuran (45 mL), and hydrochloricsolution (15 mL of a 3M solution in tetrahydrofuran). The resultingsolution was stirred for 24 h at room temperature and then thetetrahydrofuran was removed under vacuum. The resulting solution wasextracted with 3×50 mL of ethyl acetate and the combined organic layerswas washed with 1×25 mL of Na₂CO₃ (sat. aq.), dried over anhydroussodium sulfate and concentrated under vacuum to obtain 1.4 g (87%) of3,3-dimethyl-1-oxaspiro[4.5]decan-8-one as light yellow oil. ¹H NMR (300MHz, CDCl₃): 63.58 (s, 2H), 2.78-2.60 (m, 2H), 2.32-2.17 (m, 2H),2.17-2.05 (m, 2H), 1.92-1.75 (m, 2H), 1.88 (s, 2H) 1.15 (s, 6H) ppm.

Step 7: 3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yltrifluoromethanesulfonate

Into a 250-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed LiHMDS (12 mL of a 1 M solution intetrahydrofuran). A solution of 3,3-dimethyl-1-oxaspiro[4.5]decan-8-one(1.46 g, 8.01 mmol) in tetrahydrofuran (10 mL) was added at −50° C. andthe reaction mixture stirred at −50° C. for 15 min. To this was added asolution of1,1,1-trifluoro-N-phenyl-N-(trifluoromethane)sulfonylmethanesulfonamide(2.86 g, 8.01 mmol) in tetrahydrofuran (30 mL) at −50° C. The resultingsolution was stirred for 1 h at −50° C. and then for an additional 1 hat room temperature. The resulting mixture was concentrated undervacuum. The residue was purified by silica gel column with ethylacetate/petroleum ether (1:9) to obtain 1.23 g (49%) of3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate aslight yellow oil. ¹H NMR (300 MHz, CDCl₃): δ5.64 (d, J=2.7 Hz, 1H),3.57-3.50 (m, 2H), 2.69-2.50 (m, 1H), 2.50-2.22 (m, 3H), 2.01-1.87 (m,1H), 1.85-1.72 (m, 1H), 1.72-1.51 (m, 2H), 1.12 (s, 6H) ppm.

Step 8:2-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolanepin

Into a 50-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate(1.26 g, 4.01 mmol),4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(1.22 g, 4.80 mmol), 1,4-dioxane (15 mL), potassium acetate (1.18 g,12.02 mmol), and PdCl₂(dppf)CH₂Cl₂ (327 mg, 0.40 mmol). The resultingsolution was stirred for 15 h at 100° C. and then concentrated undervacuum. The residue was purified by silica gel column with ethylacetate/petroleum ether (10:1) to obtain 0.97 g (83%) of2-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolaneas yellow oil. ¹H NMR (400 MHz, CDCl₃): δ6.46 (d, J=1.6 Hz, 1H), 3.52(s, 2H), 2.50-2.07 (m, 4H), 1.80-1.54 (m, 4H), 1.26 (s, 12H), 1.11 (s,6H) ppm.

Step 9:3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde

Into a 20-mL sealed tube purged and maintained with an inert atmosphereof nitrogen, was placed 3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde(964 mg, 3.15 mmol),2-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(920 mg, 3.15 mmol), Cs₂CO₃ (3080 mg, 9.45 mmol), 1,4-dioxane/H₂O(v/v=10:1) (10 mL), and PdCl₂(dppf)CH₂Cl₂ (257 mg, 0.31 mmol). Theresulting solution was stirred for 15 h at 100° C. and then concentratedunder vacuum. The resulting residue was diluted with 50 mL of H₂O andthen the mixture was extracted with 2×30 mL of ethyl acetate and theorganic layers were combined and dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was purified by silica gel columnwith ethyl acetate/petroleum ether (1:2) to obtain 630 mg (58%) of3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehydeas yellow oil. LCMS: m/z=345.2[M+1]; ¹H NMR (400 MHz, CDCl₃): δ9.90 (s,1H), 8.13 (s, 1H), 6.30-6.20 (m, 1H), 5.40-5.30 (m, 1H), 4.15-4.00 (m,1H), 3.78-3.64 (m, 1H), 3.57 (s, 2H), 2.86-2.30 (m, 4H), 2.20-1.86 (m,4H), 1.86-1.60 (m, 6H), 1.13 (s, 6H) ppm.

Step 10: tert-butylN-(2-[[(3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

Into a 100-mL round-bottom flask, was placed3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde(630 mg, 1.83 mmol), tert-butylN-methyl-N-[2-(methylamino)ethyl]carbamate (516 mg, 2.74 mmol),ClCH₂CH₂Cl (20 mL), and NaBH(AcO)₃ (3.1 g, 14.62 mmol). The resultingsolution was stirred for 5 h at 0° C. and then quenched by the additionof 30 mL of Na₂CO₃ (sat. aq.). The resulting solution was extracted with3×50 mL of ethyl acetate and the organic layers combined and dried overanhydrous sodium sulfate and concentrated under vacuum. The residue waspurified by silica gel column with ethyl acetate/petroleum ether (3:2)to obtain 720 mg (76%) of tert-butylN-(2-[[(3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas light yellow oil. LCMS: m/z=517.5 [M+1]; ¹H NMR (400 MHz, CDCl₃):δ7.46 (s, 1H), 6.07 (s, 1H), 5.45-5.35 (m, 1H), 4.10-4.00 (m, 1H),3.72-3.62 (m, 1H), 3.60-3.50 (m, 2H), 3.45-3.20 (m, 4H), 2.83 (s, 3H),2.77-2.64 (m, 1H), 2.64-2.28 (m, 5H), 2.22 (s, 3H), 2.13-1.96 (m, 3H),1.90-1.52 (m, 7H), 1.32 (s, 9H), 1.13 (s, 3H), 1.11 (s, 3H) ppm.

Step 11: tert-butylN-(2-[[(3-[3,3-dimethyl-2-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]-ethyl)-N-methylcarbamate

Into a 30-mL pressure tank reactor, was placed tert-butylN-(2-[[(3-[3,3-dimethyl-2-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(720 mg, 1.39 mmol), acetic acid (10 mL), and 10% palladium/carbon (100mg). The reaction mixture was then subjected to hydrogen gas (pressure10 atm) and stirred for 6 h at 50° C. The solids were removed byfiltration and the solution was concentrated under vacuum to obtain 1 g(97%) of tert-butylN-(2-[[(3-[3,3-dimethyl-2-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]-ethyl)-N-methylcarbamateas light yellow oil.

Step 12:Methyl[2-(methylamino)ethyl]([3-[(5s,8s)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amineandmethyl[2-(methylamino)ethyl]([3-[(5r,8r)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amine(Compound 134 & Compound 135)

Into a 50-mL round-bottom flask, was placed tert-butylN-(2-[[(3-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(1 g, 1.93 mmol), and hydrogen chloride/methanol (saturated, 10 mL). Theresulting solution was stirred for 5 h at room temperature and thenconcentrated under vacuum. The residue was purified by Prep-HPLC withthe following conditions (Prep-HPLC-045): Column, Jupiter 4u Proteo 90A,AXIA Packed, 21.2×250 mm 4 um 9 nm; mobile phase, water with 0.05% TFAand ACN (5.0% ACN up to 30.0% in 8 min, hold 30.0% in 2 min); Detector,UV 220 nm. This resulted in 480.8 mg (44%) ofmethyl[2-(methylamino)ethyl]([3-[(5 s,8s)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)aminetrifluoroacetate salt as a white semi-solid; LCMS: m/z=335.2 [M+1]; ¹HNMR (300 MHz, D₂O): δ7.73 (s, 1H), 4.28 (s, 2H), 3.50-3.40 (m, 6H),2.74-2.68 (m, 7H), 1.90-1.86 (m, 2H), 1.68-1.42 (m, 8H) 1.00 (s, 6H)ppm. And 152.6 mg (14%) ofmethyl[2-(methylamino)ethyl]([3-[(5r,8r)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)aminetrifluoroacetate salt as a white semi-solid; LCMS: m/z=335.2 [M+1];]; ¹HNMR (300 MHz, D₂O): δ7.73 (s, 1H), 4.28 (s, 2H), 3.46-3.34 (m, 6H),2.74-2.69 (m, 7H), 1.83-1.75 (m, 4H), 1.70 (m, 2H), 1.57-1.46 (m, 4H),1.00 (s, 6H) ppm.

Compound 158([3-[4,4-bis(ethoxymethyl)cyclohexyl]-1H-pyrazol-4-yl]methyl)(methyl)[2-(methylamino)ethyl]aminehydrochloride salt

Step 1: ethyl 1,4-dioxaspiro[4.5]decane-8-carboxylate

Into a 500-mL round-bottom flask was placed ethyl4-oxocyclohexane-1-carboxylate (150 g, 881.29 mmol, 1.00 equiv),cyclohexane (300 mL), H₂NSO₃H (3 g) and ethane-1,2-diol (65.7 g, 1.06mol, 1.20 equiv). The resulting solution was stirred overnight at 100°C. and then diluted with 300 mL of ethyl acetate. The resulting mixturewas washed with 2×200 mL of brine and then concentrated under vacuum.This resulted in 152 g (80%) of ethyl1,4-dioxaspiro[4.5]decane-8-carboxylate as yellow oil. ¹H-NMR (300 MHz,CDCl₃): δ 4.05 (q, J=7.1 Hz, 2H), 3.95 (s, 4H), 2.44-2.23 (m, 1H),2.00-1.70 (m, 6H), 1.65-1.47 (m, 2H), 1.25 (t, J=7.1 Hz, 3H) ppm.

Step 2: 8,8-diethyl 1,4-dioxaspiro[4.5]decane-8,8-dicarboxylate

Into a 2-L 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen was placed (i-Pr)2NH (45.2 g) andtetrahydrofuran (1500 mL). Then n-BuLi (2.5M in hexane, 179.8 mL) wasadded dropwise at −50° C. The resulting mixture was reacted for 30 minat −50° C. Then ethyl 1,4-dioxaspiro[4.5]decane-8-carboxylate (80 g,373.38 mmol, 1.00 equiv) was added into mixture at −78° C. After 1 hour,chloro(ethoxy)methanone (60 g, 552.87 mmol, 1.48 equiv) was addeddropwise at −78° C. The resulting solution was stirred for another 30min at −78° C. The reaction was then quenched by the addition of 500 mLof water. The resulting solution was extracted with 3×800 mL of ethylacetate and the organic layers combined and concentrated under vacuum.This resulted in 82 g (77%) of 8,8-diethyl1,4-dioxaspiro[4.5]decane-8,8-dicarboxylate as light yellow oil. ¹H-NMR(400 MHz, CDCl₃): δ 4.18 (q, J=7.1 Hz, 4H), 3.94 (s, 4H), 2.18 (t, J=6.2Hz, 4H), 1.69 (t, J=6.2 Hz, 4H), 1.25 (t, J=7.1 Hz, 6H) ppm.

Step 3: [8-(hydroxymethyl)-1,4-dioxaspiro[4.5]decan-8-yl]methanol

Into a 2500-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen was placed LiAlH₄ (47.9 g, 1.26 mol, 4.02equiv) and tetrahydrofuran (1 L). This was followed by the addition of8,8-diethyl 1,4-dioxaspiro[4.5]decane-8,8-dicarboxylate (90 g, 314.33mmol, 1.00 equiv) in tetrahydrofuran (200 mL) dropwise with stirring at−20° C. The resulting solution was stirred overnight at roomtemperature. The reaction was then quenched by the addition of 500 g ofNa₂SO₄.10H₂O. The solids were filtered out. The resulting mixture wasconcentrated under vacuum to give 35 g (55%) of[8-(hydroxymethyl)-1,4-dioxaspiro[4.5]decan-8-yl]methanol as a whitesolid. ¹H-NMR (300 MHz, MeOD): δ 3.94 (s, 4H), 3.49 (s, 4H), 1.69-1.59(m, 4H), 1.59-1.44 (m, 4H) ppm.

Step 4: 8,8-bis(ethoxymethyl)-1,4-dioxaspiro[4.5]decane

Into a 1000-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen was placed[8-(hydroxymethyl)-1,4-dioxaspiro[4.5]decan-8-yl]methanol (35 g, 173.06mmol, 1.00 equiv) and N,N-dimethylformamide (400 mL). This was followedby the addition of sodium hydride (21 g, 525.00 mmol, 3.03 equiv, 60%),in portions at 0° C. The mixture was stirred for 30 min at roomtemperature. To this was added iodoethane (108 g, 692.46 mmol, 4.00equiv) dropwise with stirring. The resulting solution was stirredovernight at room temperature. The reaction was then quenched by theaddition of 300 mL of water. The resulting solution was extracted with3×200 mL of ethyl acetate and the organic layers combined. The resultingmixture was washed with 300 mL of brine. The mixture was dried overanhydrous sodium sulfate and concentrated under vacuum. The residue waspurified by silica gel column with ethyl acetate/petroleum ether (1:20).This resulted in 30 g (67%) of8,8-bis(ethoxymethyl)-1,4-dioxaspiro[4.5]decane as yellow oil. ¹H-NMR(300 MHz, CDCl₃) δ: 3.93 (s, 4H), 3.46 (q, J=7.0 Hz, 4H), 3.29 (s, 4H),1.65-1.50 (m, 8H), 1.16 (t, J=7.0 Hz, 6H) ppm.

Step 5: 4,4-bis(ethoxymethyl)cyclohexan-1-one

Into a 1000-mL round-bottom flask, was placed8,8-bis(ethoxymethyl)-1,4-dioxaspiro[4.5]decane (30 g, 116.12 mmol, 1.00equiv), FeCl₃-6H₂O (62 g, 230.48 mmol, 1.98 equiv) and dichloromethane(300 mL). The resulting solution was stirred overnight at roomtemperature. The resulting mixture was washed with 3×150 mL of water and150 mL of Na₂CO₃ (sat. aq.). The resulting mixture was washed with 150mL of brine. The mixture was dried over anhydrous sodium sulfate andconcentrated under vacuum to give 22.8 g (92%) of4,4-bis(ethoxymethyl)cyclohexan-1-one as yellow oil. ¹H-NMR (300 MHz,CDCl₃) δ: 3.46 (q, J=7.0 Hz, 4H), 3.37 (s, 4H), 2.36 (t, J=14.1 Hz, 4H),1.77 (t, J=14.1 Hz, 4H), 1.18 (t, J=7.0 Hz, 6H) ppm.

Step 6: 4,4-bis(ethoxymethyl)cyclohex-1-en-1-yltrifluoromethanesulfonate

Into a 1-L 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen was placed4,4-bis(ethoxymethyl)cyclohexan-1-one (22.8 g, 106.39 mmol, 1.00 equiv)and THF (400 mL). This was followed by the addition of LiHMDS (1M inTHF, 117.2 mL) dropwise with stirring at −50° C. The resulting solutionwas stirred for 1 hr. at −30° C. To this was added a solution of1,1,1-trifluoro-N-phenyl-N-(trifluoromethane)sulfonylmethanesulfonamide(41.8 g, 117.00 mmol, 1.10 equiv) in tetrahydrofuran (40 mL) dropwisewith stirring at −30° C. The resulting solution was allowed to react,with stirring, for an additional 4 hrs. at room temperature. Thereaction was then quenched by the addition of 100 mL of NH₄Cl (sat.aq.). The resulting solution was extracted with 2×100 mL of ethylacetate and the organic layers combined. The resulting mixture wasconcentrated under vacuum. The residue was purified by silica gel columnwith petroleum ether (100%) to give 29 g (79%) of4,4-bis(ethoxymethyl)cyclohex-1-en-1-yl trifluoromethanesulfonate asbrown oil. ¹H-NMR (300 MHz, CDCl₃) δ: 5.78-5.61 (m, 1H), 3.44 (q, J=7.0Hz, 4H), 3.27 (q, J=7.0 Hz, 4H), 2.49-2.21 (m, 2H), 2.20-2.00 (m, 2H),1.74 (t, J=6.5 Hz, 2H), 1.18 (t, J=7.0 Hz, 6H) ppm.

Step 7:2-[4,4-bis(ethoxymethyl)cyclohex-1-en-1-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

Into a 100-mL round-bottom flask purged and maintained with an inertatmosphere of nitrogen, was placed4,4-bis(ethoxymethyl)cyclohex-1-en-1-yl trifluoromethanesulfonate (29 g,83.8 mmol, 1.00 equiv), KOAc (32.4 g, 331 mmol, 3.95 equiv), Pd(dppf)Cl₂(6.13 g, 8.38 mmol, 0.10 equiv),4,4,5,5-tetramethyl-2-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(25.5 g, 100.6 mmol, 1.19 equiv) and 1,4-dioxane (300 mL). The resultingsolution was stirred overnight at 100° C. in an oil bath. The reactionwas then quenched by the addition of 200 mL of water. The resultingsolution was extracted with 3×100 mL of ethyl acetate and the organiclayers combined and dried over anhydrous sodium sulfate. The solids werefiltered out and the solution concentrated under vacuum. The residue waspurified by silica gel column with ethyl acetate/petroleum ether (1:20).This resulted in 22 g (81%) of2-[4,4-bis(ethoxymethyl)cyclohex-1-en-1-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolaneas colorless oil. ¹H-NMR (300 MHz, CDCl₃) δ: 6.67-6.35 (m, 1H), 3.44 (q,J=7.0 Hz, 4H), 3.24 (q, J=7.0 Hz, 4H), 2.18-2.05 (m, 2H), 2.03-1.84 (m,2H), 1.50 (t, J=6.3 Hz, 2H), 1.15 (t, J=7.0 Hz, 6H) ppm.

Step 8: tert-butylN-[2-[([3-[4,4-bis(ethoxymethyl)cyclohex-1-en-1-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl)(methyl)amino]ethyl]-N-methylcarbamate

Into a 1-L round-bottom flask purged and maintained with an inertatmosphere of nitrogen was placed2-[4,4-bis(ethoxymethyl)cyclohex-1-en-1-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(22 g, 67.85 mmol, 1.00 equiv), Pd(dppf)Cl₂ (3.38 g, 4.62 mmol, 0.07equiv), potassium carbonate (19.2 g, 138.92 mmol, 2.05 equiv), water (50mL), tert-butylN-[2-([3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl(methyl)amino)ethyl]-N-methylcarbamate(18 g, 37.63 mmol, 0.55 equiv) and 1,4-dioxane (500 mL). The resultingsolution was stirred overnight at 100° C. in an oil bath. The solidswere filtered out. The resulting mixture was concentrated under vacuum.The residue was purified by silica gel column with ethylacetate/petroleum ether (50%). This resulted in 18 g (48%) of tert-butylN-[2-[([3-[4,4-bis(ethoxymethyl)cyclohex-1-en-1-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl)(methyl)amino]ethyl]-N-methylcarbamateas brown oil. ¹H-NMR (300 MHz, CDCl₃) δ: 7.46 (s, 1H), 6.08 (s, 1H),5.40-5.22 (m, 1H), 4.12-4.00 (m, 1H), 3.76-3.60 (m, 1H), 3.58-3.20 (m,8H), 2.83 (s, 3H), 2.57 (s, 3H), 2.45 (s, 2H), 2.15-1.95 (m, 4H),1.82-1.52 (m, 6H), 1.44 (s, 6H), 1.35 (s, 9H), 1.15 (t, J=7.0 Hz, 6H)ppm.

Step 9: tert-butylN-[2-[([3-[4,4-bis(ethoxymethyl)cyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl)(methyl)amino]ethyl]-N-methylcarbamate

Into a 1-L round-bottom flask, was placed tert-butylN-[2-[([3-[4,4-bis(ethoxymethyl)cyclohex-1-en-1-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl)(methyl)amino]ethyl]-N-methylcarbamate(18.0 g, 32.85 mmol, 1.00 equiv), 10% Pd(OH)₂/C (20 g) andtetrahydrofuran (400 mL). Hydrogen (3 atm) was then applied to thereaction mixture. The resulting solution was stirred for 7 h at roomtemperature. The solids were filtered out and the solution concentratedunder vacuum. The residue was purified by silica gel column withdichloromethane/methanol (3.5%). This resulted in 8.8 g (49%) oftert-butylN-[2-[([3-[4,4-bis(ethoxymethyl)cyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl)(methyl)amino]ethyl]-N-methylcarbamateas yellow oil. ¹H-NMR (400 MHz, CDCl₃) δ: 7.36 (s, 1H), 5.30-5.10 (m,1H), 4.00-3.85 (m, 1H), 3.68-3.50 (m, 2H), 3.56-3.46 (m, 6H), 3.35-3.27(m, 4H), 3.14 (s, 2H), 2.77 (s, 3H), 2.69-2.37 (m, 3H), 1.94 (s, 3H),1.80-1.46 (m, 9H), 1.37 (s, 9H), 1.30-1.15 (m, 4H), 1.10 (t, J=7.0 Hz,6H) ppm.

Step 10:([3-[4,4-bis(ethoxymethyl)cyclohexyl]-1H-pyrazol-4-yl]methyl)(methyl)[2-(methylamino)ethyl]aminehydrochloride salt (Compound 158)

Into a 500-mL round-bottom flask was placed tert-butylN-[2-[([3-[4,4-bis(ethoxymethyl)cyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl)(methyl)amino]ethyl]-N-methylcarbamate(8.8 g, 15.98 mmol, 1.00 equiv) and dichloromethane (300 mL). Hydrogenchloride gas was bubbled into the reaction mixture. The resultingsolution was stirred for 4 h at room temperature and then concentratedunder vacuum. The resulting residue was washed with 1 L of hexane. Thesolids were collected by filtration. This resulted in 5.90 g (84%) of([3-[4,4-bis(ethoxymethyl)cyclohexyl]-1H-pyrazol-4-yl]methyl)(methyl)[2-(methylamino)ethyl]aminehydrochloride salt as an off-white solid. ¹H-NMR (300 MHz, D₂O) δ: 7.75(s, 1H), 4.30 (s, 2H), 3.57-3.43 (m, 10H), 3.23 (s, 2H), 2.80-2.67 (m,7H), 1.64-1.54 (m, 6H), 1.35-1.20 (m, 2H), 1.15-1.05 (m, 6H) ppm. LCMS(method M, ESI), RT=1.25 min, m/z=367.3 [M-2HCl+H]⁺.

Compound 1829-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]spiro[4.5]decan-6-ol

Step 1: 1,4-dioxadispiro[4.1.4⁷.3⁵]tetradecan-12-one

To a solution of 1,4-dioxaspiro[4.5]decan-8-one (5 g, 32.01 mmol) intoluene (90 ml) was added tBuOK (632.26 mg, 5.63 mmol) and the reactionmixture stirred at room temperature for 5 mins. 1,4-Dibromobutane (4 ml,33.62 mmol) was added and the reaction was heated at reflux for 20hours. The reaction was monitored by TLC (heptane:EtOAc 80/20, PMA). Thereaction was cooled at room temperature, quenched with an aqueoussaturated NH₄Cl solution and then diluted with EtOAc. The two layerswere separated and the aqueous layer was further extracted with EtOAc(1×). The combined organic layers were washed with water (1×) and brine(1×), dried (MgSO₄) and concentrated to give a crude oil. This productwas dissolved in a minimum amount of DCM and loaded on a 340 g SNAP KPcolumn and eluted with heptane:EtOAc 6% to 40% on Biotage to give: 3.9 g(58%) of 1,4-dioxadispiro[4.1.4⁷.3⁵] tetradecan-12-one. ¹H-NMR (500 MHz,Chloroform-d) δ 4.02-3.88 (m, 4H), 2.59-2.50 (m, 2H), 2.05 (dd, J=7.9,4.7 Hz, 2H), 1.99-1.95 (m, 2H), 1.93 (s, 2H), 1.60-1.46 (m, 6H).

Step 2: 1,4-dioxadispiro[4.1.4⁷.3⁵]tetradecan-12-ol

A solution of 1,4-dioxadispiro[4.1.4⁷.3⁵] tetradecan-12-one (3.9 g,18.55 mmol) in MeOH (100 ml) was cooled to 0° C. NaBH₄ (1.75 g, 46.37mmol) was added in small portions. The reaction was left to stir at 0°C. for 1 hour and then quenched with water. The resulting mixture wasstirred for 10 mins at room temperature and then diluted with water andEtOAc. The aqueous layer was back extracted with EtOAc and the combinedorganic layers were washed with water (1×) and brine (2×), dried (MgSO₄)filtered and the filtrate was concentrated in vacuo to give an oil, 2.96g (75%) of 1,4-dioxadispiro[4.1.4⁷.3⁵]tetradecan-12-ol. This materialwas used in the next step without further purification. ¹H-NMR (500 MHz,Chloroform-d) δ 3.93 (s, 4H), 3.52 (dd, J=6.7, 2.4 Hz, 1H), 1.90-1.76(m, 3H), 1.77-1.42 (m, 12H).

Step 3:tert-butyl({1,4-dioxadispiro[4.1.4⁷.3⁵]tetradecan-12-yloxy})dimethylsilane

To a solution of 1,4-dioxadispiro[4.1.4⁷.3⁵]tetradecan-12-ol (2.96 g,13.94 mmol) in DMF (40 ml) was added tert-butyl(chloro)dimethylsilane(2.31 g, 15.34 mmol) and 1H-imidazole (1.9 g, 27.89 mmol). The reactionwas stirred at room temperature overnight. The reaction mixture wasdiluted with water and extracted with ether (3×). The combined organiclayers were washed with water (2×) and brine (2×), dried (MgSO₄),filtered and concentrated to give an oil. This was purified bydissolving in a minimum amount of DCM, loading on a 25 g KP SNAP column,eluting with 5%-35% of EtOAc in heptane to give 3.73 g (82%) oftert-butyl({1,4-dioxadispiro[4.1.4⁷.3⁵]tetradecan-12-yloxy})dimethylsilane. ¹H-NMR (500 MHz, Chloroform-d) δ 3.97-3.83 (m, 4H), 3.44(dd, J=5.5, 2.2 Hz, 1H), 1.89 (td, J=12.1, 4.0 Hz, 1H), 1.83 (d, J=13.5Hz, 1H), 1.79-1.46 (m, 9H), 1.43-1.30 (m, 3H), 0.89 (s, 9H), 0.04 (s,6H).

Step 4: 10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]decan-7-one

To a solution oftert-butyl({1,4-dioxadispiro[4.1.4⁷.3⁵]tetradecan-12-yloxy})dimethylsilane(3.7 g, 11.33 mmol) in DCM (120 ml) was added trichloroiron hexahydrate(15.31 g, 56.65 mmol) and the resulting suspension was stirred at roomtemperature for 2 hours. The reaction was monitored by TLC (9:1heptane/EtOAc, DNP stain). The reaction mixture was diluted with DCM,decanted to remove most of the solid inorganics and washed with aqueoussaturated NaHCO₃ (1×), water (1×), brine (1×), dried over MgSO₄,filtered and the filtrate was concentrated in vacuo to give a clear oil:3.22 g (99%) of 10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]decan-7-one.This material was used in the next step. ¹H-NMR (500 MHz, Chloroform-d)δ 3.62 (t, J=3.3 Hz, 1H), 2.67-2.54 (m, 2H), 2.16 (dtd, J=14.2, 4.6, 2.1Hz, 1H), 2.06 (d, J=13.4 Hz, 1H), 1.90 (dt, J=8.1, 4.2 Hz, 2H),1.76-1.48 (m, 5H), 1.38 (ddd, J=12.1, 6.7, 5.2 Hz, 1H), 1.28 (ddd,J=19.8, 7.7, 4.8 Hz, 2H), 0.92 (s, 9H), 0.10 (d, J=2.9 Hz, 6H).

Step 5: 10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-6-en-7-yltrifluoromethane sulfonate and10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-7-en-7-yltrifluoromethane sulfonate

A solution of 10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]decan-7-one(3.2 g, 11.33 mmol) in THF (45 ml) was cooled to −78° C. under nitrogen.A 2M solution of LDA in heptanes-THF (7.9 ml) was added and theresulting solution was stirred at −78° C. for 1 hour.1,1,1-Trifluoro-N-phenyl-N-[(trifluoromethyl)sulfonyl]methanesulfonamide(4.45 g, 12.46 mmol) was added at −78° C. to the reaction as a solutionin THF (15 ml) and the reaction was stirred at −78° C. for 1 hour andthen left to warm to room temperature and stir overnight. The reactionwas quenched by addition of water and diluted with EtOAc. The combinedorganic layers were washed with water (1×) and brine (2×), dried(MgSO₄), filtered and the filtrate was concentrated in vacuo to give anoil. This material was dissolved in a minimum amount of DCM and wasloaded on a 100 g KP SNAP column and eluted with heptane-EtOAc 0% to25%, to give 2.81 g (60%) of a mixture of10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-6-en-7-yltrifluoromethanesulfonate and10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-7-en-7-yltrifluoromethanesulfonate as a clear oil. NMR showed a ratio of 9:1 forthe two isomers. ¹H-NMR (500 MHz, Chloroform-d) δ 5.59 (t, J=3.8 Hz,0.9H), 5.53 (s, 0.1H), 3.60 (dd, J=6.2, 3.3 Hz, 0.1H), 3.56 (t, J=4.2Hz, 0.9H), 2.46-2.40 (m, 1H), 2.40-2.32 (m, 1H), 2.13 (ddq, J=17.9, 4.2,2.2 Hz, 1H), 2.07 (d, J=16.6 Hz, 1H), 1.70-1.57 (m, 5H), 1.46-1.32 (m,3H), 0.87 (s, 9H), 0.06 (d, J=4.3 Hz, 6H).

Step 6:tert-butyldimethyl{[9-(tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[4.5]dec-8-en-6-yl]oxy}silaneandtert-butyldimethyl{[9-(tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[4.5]dec-9-en-6-yl]oxy}silane

A suspension of 10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-6-en-7-yltrifluoromethanesulfonate and10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-7-en-7-yltrifluoromethanesulfonate (0.74 g, 2.89 mmol), potassium acetate (1.78g, 18.09 mmol), bis[3-(diphenylphosphanyl)cyclopenta-2,4-dien-1-yl]iron;dichloromethane, dichloropalladium (99 mg, 0.12 mmol) in 1,4-dioxane (10ml) was purged with nitrogen for 5 minutes and then heated at 80° C. ina pressure tube overnight. The mixture was allowed to cool at roomtemperature and diluted with EtOAc and filtered through Celite®. Thefiltrate was washed with water (1×), brine (1×) and dried over MgSO₄.The solvent was evaporated and the residue purified on a 50 g KP SNAPcolumn on Biotage eluting with a gradient of heptane:EtOAc (0% to 20%)to give 0.41 g (43%) of a mixture oftert-butyldimethyl{[9-(tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[4.5]dec-8-en-6-yl]oxy}silaneandtert-butyldimethyl{[9-(tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[4.5]dec-9-en-6-yl]oxy}silane,409 mg (43%) in a 9:1 ratio as shown by NMR. ¹H-NMR (500 MHz,Chloroform-d) δ 6.45-6.38 (m, 0.9H), 6.28 (t, J=1.8 Hz, 0.1H), 3.67-3.57(m, 1H), 2.29 (dtt, J=18.7, 4.2, 2.0 Hz, 1H), 2.20 (dd, J=17.4, 1.7 Hz,1H), 2.12-2.01 (m, 1H), 1.94 (dt, J=17.5, 2.5 Hz, 1H), 1.71-1.45 (m,8H), 1.26 (s, 12H), 0.87 (s, 9H), 0.02 (d, J=3.7 Hz, 6H).

Step 7A: tert-butylN-[2-({[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl}(methyl)amino)ethyl]-N-methylcarbamate

To a solution of 3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde (7.42g, 24.25 mmol) in 1,2-dichloroethane (160 ml) was added tert-butylN-methyl-N-[2-(methylamino)ethyl]carbamate (90%, 6.14 g, 29.34 mmol)followed by the addition of NaBH(OAc)₃ (10.28 g, 48.49 mmol). Theresulting mixture was stirred at room temperature. The reaction mixturewas diluted with DCM, washed with brine (2×), dried over MgSO₄, anddried in vacuo. Purification by flash chromatography using a BiotageIsolera system with a 100 g KP SNAP cartridge, eluting with a gradientof MeOH in DCM (0 to 10%,) afforded tert-butylN-[2-({[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl}(methyl)amino)ethyl]-N-methylcarbamate as a white solid after crystallization from heptane-ether 8.4g (72%). ¹H NMR (500 MHz, Chloroform-d) δ 7.88-7.35 (m, 1H), 5.32 (dd,J=9.5, 2.6 Hz, 1H), 4.09-3.99 (m, 1H), 3.72-3.58 (m, 1H), 3.55-3.23 (m,4H), 2.85 (s, 3H), 2.69-2.43 (m, 2H), 2.39-2.16 (m, 3H), 2.14-1.92 (m,3H), 1.73-1.49 (m, 3H), 1.44 (s, 9H).

Step 7B: tert-butylN-(2-{[(3-{10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-7-en-7-yl}-1-(oxan-2-yl)pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate

A suspension of tert-butylN-[2-({[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl}(methyl)amino)ethyl]-N-methylcarbamate(488 mg, 1.02 mmol),tert-butyldimethyl{[9-(tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[4.5]dec-8-en-6-yl]oxy}silaneandtert-butyldimethyl{[9-(tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[4.5]dec-9-en-6-yl]oxy}silane(400 mg, 1.02 mmol) in 1,4-dioxane (2 ml) and aqueous 2M sodiumcarbonate (1.53 ml) was degassed by bubbling nitrogen for 5 mins.Bis[3-(diphenylphosphanyl)cyclopenta-2,4-dien-1-yl]iron,dichloromethane, and dichloropalladium (42 mg, 0.05 mmol) was added andthe reaction was heated at 80° C. overnight. The reaction mixture wasleft to cool to room temperature and then diluted with EtOAc, filteredthrough Celite® and the solids were washed with EtOAc. The combinedfiltrates were washed with water (1×) and brine (2×), dried (MgSO₄),filtered and concentrated to give an orange oil. This material waspurified by loading as a solution in DCM on a 25 g SNAP column onBiotage and elution with a gradient of heptane-EtOAc 35% to 100% to givetert-butylN-(2-{[(3-{10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-7-en-7-yl}-1-(oxan-2-yl)pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate,0.45 g (72%). LC-MS: 2.42 min (3 min method), m/z=617.35. ¹H-NMR (500MHz, Chloroform-d) δ 7.41 (d, J=3.9 Hz, 1H), 5.91 (d, J=21.6 Hz, 1H),5.26 (dd, J=8.9, 2.8 Hz, 1H), 4.04-3.97 (m, 1H), 3.69-3.56 (m, 2H), 3.27(d, J=49.4 Hz, 4H), 2.77 (s, 3H), 2.53-2.37 (m, 3H), 2.33 (d, J=18.3 Hz,1H), 2.25 (d, J=16.6 Hz, 1H), 2.17 (s, 3H), 2.14-2.05 (m, 1H), 1.98 (dd,J=10.2, 2.9 Hz, 3H), 1.67-1.46 (m, 11H), 1.38 (s, 9H), 0.84-0.80 (m,9H), 0.00 (s, 6H).

Step 8: tert-butylN-(2-{[(3-{10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]decan-7-yl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate

A suspension of tert-butylN-(2-{[(3-{10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]dec-7-en-7-yl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate(453 mg, 0.73 mmol) and Pd/C (10%, 156 mg, 0.147 mmol) in EtOH (10 ml)was stirred at room temperature under an atmosphere of hydrogen for 2days. The reaction mixture was filtered through Celite® and the filtrateconcentrated to give an oil that was purified on Biotage using a SNAP KP25 g column, eluting with a gradient of heptane-EtOAc 25% to 100% togive tert-butylN-(2-{[(3-{10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]decan-7-yl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate,120 mg (26%). LC-MS: 2.16 min (3 min method), m/z=619.35. ¹H-NMR (500MHz, Chloroform-d) δ 7.46 (d, J=10.4 Hz, 1H), 5.31-5.24 (m, 1H), 4.05(d, J=11.5 Hz, 1H), 3.67 (td, J=11.2, 2.2 Hz, 1H), 3.48-3.21 (m, 5H),2.88-2.73 (m, 4H), 2.55-2.38 (m, 2H), 2.22 (s, 3H), 2.07-1.88 (m, 6H),1.70-1.48 (m, 12H), 1.43 (d, J=10.5 Hz, 11H), 0.92 (s, 9H), 0.05 (s,6H).

Step 9:9-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]spiro[4.5]decan-6-ol(Compound 182)

A suspension of tert-butylN-(2-{[(3-{10-[(tert-butyldimethylsilyl)oxy]spiro[4.5]decan-7-yl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate(77 mg, 0.12 mmol) in aqueous 6M HCl (0.56 ml) was stirred at roomtemperature overnight. It was diluted with water and extracted with DCM(2×). The combined organic layers were concentrated to give an oilyresidue that was dissolved in 1 ml DMSO-CH₃CN (1:1) and purified on theGilson3 using a high pH prep-HPLC method to give9-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]spiro[4.5]decan-6-ol,13 mg (33%) as a white solid. LC-MS: 4.3 min (7 min, high pH),m/z=321.3. ¹H-NMR (500 MHz, Methanol-d4) δ 7.47 (s, 0.1H), 7.42 (s,0.9H), 3.49 (s, 1H), 3.42 (s, 2H), 2.93 (tt, J=12.7, 3.4 Hz, 1H), 2.71(t, J=6.5 Hz, 2H), 2.52 (t, J=6.5 Hz, 2H), 2.37 (s, 3H), 2.20 (s, 3H),1.99-1.85 (m, 2H), 1.86-1.72 (m, 3H), 1.72-1.50 (m, 6H), 1.47-1.30 (m,3H).

Compounds 183 & 184 Racemic Mixture of(1R,4S)-4-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]spiro[5.5]undecan-1-ol (Compound 183)

Racemic Mixture of(1S,4S)-4-[4-({methyl[2-(methylamino)ethyl]amino}ethyl)-1H-pyrazol-3-yl]spiro[5.5]undecan-1-ol (Compound 184)

Step 1: 1,4-dioxadispiro[4.1.5⁷.3⁵]pentadecan-13-one

To a solution of 1,4-dioxaspiro[4.5]decan-8-one (5 g, 32.01 mmol) and1,5-dibromopentane (7.36 g, 32 mmol) in toluene (120 ml) was added tBuOK(3.59 g, 32 mmol) at RT. The solution was refluxed overnight. Thereaction was cooled to RT and quenched with HCl (0.5N, 10 ml). Thephases were separated and the aqueous was extracted with DCM (3×30 ml).The organic extracts were combined and dried over Na₂SO₄ and evaporatedto dryness. The residue was purified by Biotage (SNAP 340 g, eluentheptane/EtOAc 95/5 to 60/40) to afford 2.35 g of the title compound(33%) as a light colourless oil. ¹H-NMR (500 MHz, Chloroform-d) δ4.08-3.91 (m, 4H), 2.61-2.42 (m, 2H), 2.01-1.95 (m, 2H), 1.93 (s, 2H),1.83-1.72 (m, 2H), 1.58-1.38 (m, 7H), 1.38-1.28 (m, 1H). Rf=0.47(heptane/EtOAc 7/3).

Step 2: 1,4-dioxadispiro[4.1.5⁷.3⁵]pentadecan-13-ol

Sodium borohydride (0.99 g, 26.19 mmol) was added at 0° C. and undernitrogen to 1,4-dioxadispiro[4.1.5⁷.3⁵]pentadecan-13-one (2.35 g, 10.48mmol) in MeOH (120 ml). The reaction was stirred at 0° C. untilcompletion (6 h). The reaction was quenched slowly with water (100 mL).DCM (50 mL) was added and the layers separated. The aqueous layer wasextracted with DCM (2×50 mL). The organic layers were combined, driedover Na₂SO₄, filtered and concentrated in vacuo to afford 2.18 g ofdesired 1,4-dioxadispiro[4.1.57.35]pentadecan-13-ol (92%). ¹H-NMR (500MHz, Chloroform-d) δ 3.97-3.86 (m, 4H), 3.52-3.44 (m, 1H), 1.90-1.79 (m,3H), 1.77-1.62 (m, 2H), 1.61-1.34 (m, 10H), 1.35-1.18 (m, 2H). Rf=0.35(heptane/EtOAc 7/3).

Step 3:tert-butyl({1,4-dioxadispiro[4.1.5⁷.3⁵]pentadecan-13-yloxy})dimethylsilane

To a solution of 1,4-dioxadispiro[4.1.5⁷.3⁵]pentadecan-13-ol (2.18 g,9.63 mmol) in DMF (20 ml) was added tert-butyl(chloro)dimethylsilane(2.18 g, 14.45 mmol) and 1H-imidazole (1.32 g, 19.27 mmol). The reactionwas stirred at RT overnight. The reaction mixture was diluted with waterand extracted with EtOAc (3×50 ml). The combined organic layers weredried (Na₂SO₄) and concentrated. The residue was purified by Biotage(SNAP 100 g, eluent heptane/EtOAc 95/5 to 80/20) to afford 2.45 g ofdesired material (75%). ¹H-NMR (500 MHz, Chloroform-d) δ 4.02-3.80 (m,4H), 3.49-3.38 (m, 1H), 1.89 (td, J=12.2, 4.2 Hz, 1H), 1.84-1.71 (m,2H), 1.64-1.15 (m, 13H), 0.89 (s, 9H), 0.04 (d, J=2.2 Hz, 6H). Rf=0.60(EtOAc/heptane 10/90).

Step 4: 5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undecan-2-one

To a solution oftert-butyl({1,4-dioxadispiro[4.1.5⁷.3⁵]pentadecan-13-yloxy})dimethylsilane(2.45 g, 7.19 mmol) in DCM (100 ml) was added iron trichloridehexahydrate (1.94 g, 7.19 mmol). After 2 h, no more starting materialwas detected by TLC. The reaction mixture was washed with water (50 ml),aq sat NaHCO₃ (50 ml), brine (50 ml), the organic layer was dried overNa₂SO₄ and evaporated to dryness to afford 2.08 g of ketone as a clearoil (97%). ¹H-NMR (500 MHz, Chloroform-d) δ 3.67 (s, 1H), 2.67-2.48 (m,2H), 2.26-2.11 (m, 2H), 2.00 (dddd, J=14.5, 12.2, 5.4, 2.5 Hz, 1H), 1.87(ddt, J=14.2, 7.1, 3.7 Hz, 1H), 1.64-1.17 (m, 10H), 0.92 (s, 9H), 0.10(t, J=2.9 Hz, 6H). Rf=0.40 (EtOAc/heptane 10/90).

Step 5: 5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undec-2-en-2-yltrifluoromethanesulfonate

A stirred solution of5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undecan-2-one (2 g, 6.8 mmol)was dissolved in dry THF (160 mL) and cooled to −78° C. To this 0.18MLHMDS in THF (73.5 mL) was added dropwise. The reaction was stirred for45 min and a solution ofN-(5-chloropyridin-2-yl)-1,1,1-trifluoro-N-[(trifluoromethyl)sulfonyl]methanesulfonamide (5 g, 12.75 mmol) in THF (60 mL) was added dropwise over 10minutes, the reaction was stirred at −78° C. for 1 hr, then allowed towarm to RT over 3 hr. The reaction was quenched by addition of sat.NH₄Cl (100 mL). EtOAc (100 mL) was added and the organic layerseparated. The aqueous layer was washed (2×100 mL EtOAc), and theorganics combined, dried (Na₂SO₄), filtered, and solvent removed underreduced pressure to leave 8.4 g yellow crude material. The crude productwas purified using silica gel column chromatography (Biotage SNAP-HP 100g cartridge, dry loaded, eluent heptane:EtOAc 99:1 to 9:1) to afford2.16 g of the target material as a colourless free flowing oil (71%, 95%purity). ¹H-NMR (500 MHz, Chloroform-d) δ 5.54 (d, J=4.1 Hz, 1H), 3.51(t, J=3.9 Hz, 1H), 2.48-2.30 (m, 2H), 2.16-2.06 (m, 2H), 1.64-1.17 (m,10H), 0.88 (s, 9H), 0.05 (s, 6H). Rf=0.61 (EtOAc/heptane 5/95). LC-MS:2.72 min (hydrophobic LC-MS method), no ionisation.

Step 6:tert-butyldimethyl{[4-(tetramethyl-1,3,2-dioxaborolan-2-yl)spiro[5.5]undec-3-en-1-yl]oxy}silane

5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undec-2-en-2-yltrifluoromethanesulfonate (90%, 500 mg, 1.05 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (320 mg, 1.26mmol), Pd(dppf)Cl₂ (43 mg, 0.05 mmol) and potassium acetate (770 mg,7.87 mmol) were suspended in dioxane (5 ml). The solution was degassedwith nitrogen for 10 min and then heated to 80° C. After 2 h, no SM wasvisible by LCMS but still a trace by TLC. The reaction was allowed tocool to RT and stirred O/N. Water (10 ml) was added and the reaction wasextracted with EtOAc (2×20 ml). The combined organic layers were washedwith water (10 ml) and was dried over Na₂SO₄ and evaporated to dryness.The residue was purified by Biotage (SNAP 50 g, eluent heptane/EtOAc100/0 to 90/10) to afford 310 mg of desired boronic ester (65%) as acolourless oil. ¹H-NMR (500 MHz, Chloroform-d) δ 6.57 (s, OH), 6.45-6.32(m, 1H), 3.58-3.40 (m, 1H), 2.36 (dd, J=17.6, 1.9 Hz, 1H), 2.30-2.20 (m,1H), 2.05 (ddd, J=18.8, 5.9, 3.1 Hz, 1H), 1.79 (dd, J=17.6, 2.2 Hz, 1H),1.60-1.06 (m, 22H), 0.91-0.83 (m, 9H), 0.01 (d, J=1.4 Hz, 6H). Rf=0.47(EtOAc/heptane 5/95). LC-MS: 2.85 min (hydrophobic LC-MS method), noionisation.

Step 7: tert-butylN-(2-{[(3-{5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undec-2-en-2-yl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate

tert-Butyldimethyl {[4-(tetramethyl-13,2-dioxaborolan-2-yl)spiro[5.5]undec-3-en-1-yl]oxy}silane (90%, 200 mg,0.44 mmol),tert-butyl-N-(2-{[(3-iodo-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate(140 mg, 0.29 mmol), Pd(dppf)Cl₂ (24 mg, 29 μmol) and dipotassiumcarbonate (122 mg, 0.88 mmol) were suspended in a mixture of dioxane (14ml) and water (1 ml). The reaction was degassed with nitrogen for 10 minand then heated to 100° C. under nitrogen. After overnight, TLC and LCMSshowed the presence of desired material. The solvent was removed underreduced pressure and the residue was purified by Biotage (SNAP 50 g,eluent heptane/EtOAc 83/17 to 0/100) to afford 140 mg of desired alkene(87%) as a yellow oil. ¹H-NMR (500 MHz, Chloroform-d) δ 7.56-7.40 (m,1H), 6.00-5.85 (m, 1H), 5.32 (dd, J=9.2, 2.9 Hz, 1H), 4.06 (d, J=9.9 Hz,1H), 3.72-3.64 (m, 1H), 3.60 (t, J=5.4 Hz, 1H), 3.43-3.21 (m, 3H), 2.82(s, 2H), 2.62 (d, J=14.6 Hz, 1H), 2.46 (s, 2H), 2.34 (d, J=17.8 Hz, 1H),2.19 (d, J=19.2 Hz, 4H), 1.93 (d, J=15.1 Hz, 1H), 1.76-1.34 (m, 18H),1.24 (s, 9H), 0.88 (s, 9H), 0.04 (d, J=2.3 Hz, 6H). LCMS: 1.59 min (2min method), m/z=631.25. Rf=0.30 (heptane/EtOAc, 3/7, UV and PMA).

Step 8: Racemic tert-butylN-{2-[({3-[(2S,5S)-5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undecan-2-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamateand Racemic tert-butylN-{2-[({3-[(2S,5R)-5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undecan-2-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate

tert-ButylN-(2-{[(3-{5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undec-2-en-2-yl}-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate(140 mg, 0.26 mmol) and palladium on carbon (10%) (27 mg, 0.26 mmol)were suspended in EtOH (5 ml). The reaction was stirred at RT underhydrogen atmosphere. LCMS after overnight shows only starting material.The solution was filtered on Celite and washed with MeOH (2×10 ml). Thefiltrate was evaporated and the residue was dissolved in EtOH (5 ml) andpalladium on carbon (10%) (27 mg, 0.26 mmol) added. The reaction wasstirred at RT under hydrogen atmosphere for 36 h. The solution wasfiltered on Celite and washed with MeOH (2×10 ml). The filtrate wasevaporated and the residue was purified by Biotage (SNAP HP 10 g, eluentheptane/EtOAc 95/5 to 0/100) to afford 40 mg of deaminated side product(43%) as a yellow oil, 10 mg of isomer 1 (7%) as a yellow oil and 40 mgof isomer 2 (28%) as a yellow oil. Deaminated side product: ¹H-NMR (500MHz, Chloroform-d) δ 7.30-7.27 (m, 1H), 5.31-5.23 (m, 1H), 4.10-4.00 (m,1H), 3.73-3.62 (m, 1H), 3.56-3.50 (m, 1H), 2.97-2.85 (m, 1H), 2.08 (s,3H), 2.06-1.96 (m, 4H), 1.85-1.72 (m, 2H), 1.72-1.62 (m, 4H), 1.62-1.51(m, 4H), 1.43 (dd, J=10.3, 5.5 Hz, 4H), 1.40-1.19 (m, 4H), 0.91 (d,J=3.3 Hz, 9H), 0.05 (d, J=3.9 Hz, 6H). Rf=0.80 (heptane/EtOAc 3/7).LC-MS: 2.73 min (hydrophobic LC-MS method), m/z=447.2. Isomer 1: ¹H-NMR(500 MHz, Chloroform-d) δ 7.53-7.39 (m, 1H), 5.35-5.22 (m, 1H), 4.04 (d,J=9.9 Hz, 1H), 3.75-3.61 (m, 1H), 3.56-3.21 (m, 5H), 2.96-2.78 (m, 4H),2.61-2.38 (m, 2H), 2.30-2.15 (m, 3H), 2.11-1.88 (m, 5H), 1.84-1.50 (m,12H), 1.50-1.39 (m, 14H), 1.30-1.16 (m, 13H), 0.92 (s, 9H), 0.09-−0.00(m, 6H). Rf=0.44 (heptane/EtOAc 3/7). LC-MS: 1.72 min (2.5 minute LC-MSmethod), m/z=633.25. Isomer 2: ¹H-NMR (500 MHz, Chloroform-d) δ7.56-7.37 (m, 1H), 5.32-5.24 (m, 1H), 4.04 (d, J=9.9 Hz, 1H), 3.67 (td,J=10.1, 8.9, 4.3 Hz, 1H), 3.55-3.18 (m, 5H), 2.90 (t, J=12.9 Hz, 1H),2.82 (s, 3H), 2.58-2.39 (m, 2H), 2.29-2.18 (m, 3H), 2.05-1.91 (m, 4H),1.83-1.50 (m, 9H), 1.50-1.39 (m, 13H), 1.36-1.20 (m, 5H), 0.92 (s, 9H),0.05 (d, J=4.9 Hz, 6H). Rf=0.35 (heptane/EtOAc 3/7). LC-MS: 1.66 min(2.5 minute LC-MS method), m/z=633.25.

Step 9 (Isomer 2): Racemic Mixture of(1R,4S)-4-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]spiro[5.5]undecan-1-ol(Compound 183)

tert-ButylN-(2-{[(3-{5-[(tert-butyldimethylsilyl)oxy]spiro[5.5]undecan-2-yl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate(40 mg, 0.06 mmol) was dissolved in dioxane (1 ml) and HCl (6M, 1 ml)was added. The reaction was monitored by LCMS. After 2 h, no morestarting material was left and full conversion to the desired massdetected (mass trace only). The solvent was removed under reducedpressure and the residue was purified by SCX (2 g) column eluting withMeOH (10 ml) then MeOH/NH₃ (10 ml) to afford 20 mg (85%) of the desiredmaterial at 90% purity (assessed by ¹H-NMR). ¹H-NMR (500 MHz,Chloroform-d) δ 7.40 (s, 1H), 3.67 (s, 1H), 3.36 (s, 2H), 2.99 (tt,J=13.0, 3.7 Hz, 1H), 2.68 (t, J=6.0 Hz, 2H), 2.50 (t, J=5.9 Hz, 2H),2.41 (s, 3H), 2.18-2.15 (m, 1H), 2.14 (s, 3H), 2.09-1.97 (m, 1H),1.95-1.84 (m, 1H), 1.83-1.69 (m, 3H), 1.62 (d, J=10.2 Hz, 4H), 1.54-1.26(m, 9H).

Step 9 (Isomer 1): Racemic Mixture of(1S,4S)-4-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]spiro[5.5]undecan-1-ol(Compound 184)

Similarly, 5 mg of title compound were isolated from 10 mg reaction (85%yield, 80% purity). ¹H-NMR (500 MHz, Chloroform-d) δ 7.47-7.32 (m, 1H),3.68 (s, 1H), 3.41-3.34 (m, 2H), 3.05-2.91 (m, 1H), 2.89-2.75 (m, 3H),2.64-2.55 (m, 2H), 2.53-2.44 (m, 3H), 2.31-2.20 (m, 1H), 2.19-2.14 (m,2H), 2.08-1.96 (m, 1H), 1.94-1.83 (m, 1H), 1.81-1.70 (m, 2H), 1.70-1.55(m, 4H), 1.55-1.19 (m, 10H).

Compound 1852,2-dimethyl-5-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]cyclohexan-1-ol

Step 1: 5,5-dimethyl-7-oxabicyclo[4.1.0]heptan-2-one

To an ice-cold solution of 4,4-dimethyl-cyclohex-2-enone (1.00 g, 8.05mmol) in methanol (8 mL) was added 35% hydrogen peroxide (4.6 mL, 40.91mmol) followed by 0.5 N NaOH (2.2 mL, 1.1 mmol). The mixture was stirredat 0° C. for 5 h, stored in a fridge (4° C.) overnight and then stirredat 0° C. for another 5 h. After this time the reaction mixture wasconcentrated in vacuo, then water (15 mL) was added, and the mixtureextracted with dichloromethane (3×50 mL). The organic layers werecombined, washed with 10% Na₂SO₃ (2×40 mL) and brine, dried over Na₂SO₄,filtered and concentrated at reduced pressure to afford5,5-dimethyl-7-oxa-bicyclo[4.1.0]heptan-2-one as a colourless oil (1.18g, quant): MS (ESI+) for C₈H₁₂O₂ m/z 141.0 (M+H)⁺.; HPLC purity 88%(ret. time, 1.00 min); ¹H-NMR (500 MHz, Chloroform-d) δ 3.23 (d, J=4.0Hz, 1H), 3.18 (dd, J=4.0, 1.1 Hz, 1H), 2.41 (ddd, J=18.9, 6.4, 3.0 Hz,1H), 2.19 (ddd, J=18.9, 11.7, 7.0 Hz, 1H), 1.91 (ddd, J=13.5, 11.9, 6.4Hz, 1H), 1.34 (dddd, J=13.6, 7.0, 3.0, 1.2 Hz, 1H), 1.22 (s, 3H), 1.07(s, 3H).

Step 2: 3-hydroxy-4,4-dimethylcyclohexan-1-one

Lithium (sticks) metal (74.27 mg, 10.70 mmol) was added to a solution ofnaphthalene (1.83 g, 14.27 mmol) in dry THF (25 ml) at RT and stirreduntil the metal was dissolved (˜3 h) before cooling to −78° C. Asolution of 5,5-dimethyl-7-oxabicyclo[4.1.0]heptan-2-one (0.50 g, 3.57mmol) in dry THF (10 ml) was then added and stirred for 20 min. Water (5ml) was added and the reaction was allowed to warm to RT. Water (20 ml)was added to the reaction mixture and the product extracted with Et₂O(2×50 ml). This was dried over Na₂SO₄, filtered and evaporated in vacuo.Purification by silica gel column chromatography, on a Biotage Isolerasystem, using a 25 g KP-Sil SNAP cartridge, eluting withEtOAc:heptanes+1% TEA (1:9-9:1), gave the desired product as acolourless oil (210 mg, 41%): ¹H-NMR (500 MHz, Chloroform-d) δ 3.72 (dd,J=8.1, 4.3 Hz, 1H), 2.67 (ddd, J=14.9, 4.3, 1.0 Hz, 1H), 2.48-2.40 (m,1H), 2.39-2.32 (m, 2H), 1.89 (dt, J=13.3, 6.6 Hz, 1H), 1.51 (ddd,J=14.3, 8.3, 6.4 Hz, 1H), 1.15 (s, 3H), 1.10 (s, 3H).

Step 3: 3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohexan-1-one

tert-Butyldimethylsilylchloride (0.43 g, 2.84 mmol) was added to asolution of 3-hydroxy-4,4-dimethylcyclohexan-1-one (0.34 g, 2.37 mmol)and imidazole (0.39 g, 5.69 mmol) in DCM (10 ml) at RT and stirred overthe weekend. The reaction mixture was diluted with DCM (50 ml) andwashed with water (50 ml) and then brine (50 ml), dried over Na₂SO₄,filtered and evaporated in vacuo. Purification by silica gel columnchromatography, on a Biotage Isolera system, using a 25 g KP-Sil SNAPcartridge, eluting with EtOAc:heptanes (1:9-2:8-1) gave the desiredproduct as a colourless oil (193 mg, 31%): ¹H-NMR (500 MHz,Chloroform-d) δ 3.64 (dd, J=7.4, 4.2 Hz, 1H), 2.59-2.52 (m, 1H),2.39-2.25 (m, 3H), 1.90-1.81 (m, 1H), 1.47-1.40 (m, 1H), 1.07 (s, 3H),1.01 (s, 3H), 0.88 (s, 9H), 0.05 (d, J=6.0 Hz, 6H).

Step 4: 5-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohex-1-en-1-yltrifluoromethanesulfonate

2 M LDA in THF (0.96 ml, 1.92 mmol) was added to a solution of3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohexan-1-one (0.35 g,1.37 mmol) in dry THF (10 ml) at −78° C. and stirred for 1 hr under N₂.1,1,1-trifluoro-N-phenyl-N-[(trifluoromethyl)sulfonyl]methanesulfonamide(0.59 g, 0.53 mmol) was added to the reaction as a solution in THF (1ml) and the reaction was stirred at −78° C. for 1 hour and then left towarm to RT and stirred overnight. The reaction was quenched by theaddition of water (1 ml) and diluted with ethyl acetate (30 ml). Thiswas washed with water (30 ml) then brine (30 ml), dried over Na₂SO₄,filtered and evaporated in vacuo, to give an oil. This was dissolved inminimum amount of DCM and was loaded on a 25 g KP SNAP column and elutedwith heptane-EtOAc 0% to 6% to give 335 mg (62%) of the desired productas a colourless oil: ¹H-NMR (500 MHz, Chloroform-d) δ 5.62-5.48 (m, 1H),3.54-3.44 (m, 1H), 2.50-2.42 (m, 1H), 2.24-2.15 (m, 1H), 2.11-2.03 (m,1H), 1.85-1.77 (m, 1H), 0.85-0.81 (m, 15H), -0.01 (d, J=7.3 Hz, 6H).

Step 5:tert-butyl({[6,6-dimethyl-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl]oxy})dimethylsilane

A suspension of5-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohex-1-en-1-yltrifluoromethanesulfonate (0.34 g, 0.86 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (0.33 g, 1.29mmol) and KOAc (0.59 g, 6.04 mmol) in 1,4-dioxane (5 ml) was degassedwith a nitrogen sparge for 10 min whilst stirring at RT.Bis[3-(diphenylphosphanyl)cyclopenta-2,4-dien-1-yl]iron, dichloromethaneand dichloro palladium (0.07 g, 0.09 mmol) was added to this suspensionand stirred at 90° C. for 3.5 h before allowing to cool to RT overnight.The reaction mixture was diluted with EtOAc (10 ml) and water (10 ml).The organic layer was separated and the aqueous was extracted with EtOAc(10 ml). The combined organic layers were dried over Na₂SO₄, filteredand evaporated in vacuo. Purification by silica gel columnchromatography, on a Biotage Isolera system, using a 25 g KP-Sil SNAPcartridge, eluting with EtOAc:heptanes (0-1), gave the desired productas a colourless oil (250 mg, 79%): ¹H-NMR (500 MHz, Chloroform-d) δ6.49-6.15 (m, 1H), 3.52-3.42 (m, 1H), 2.32-2.21 (m, 1H), 2.12-1.98 (m,2H), 1.93-1.83 (m, 1H), 1.25 (d, J=3.3 Hz, 12H), 0.91 (s, 3H), 0.87 (d,J=3.8 Hz, 9H), 0.82 (d, J=3.4 Hz, 3H), 0.07-−0.02 (m, 6H).

Step 6: tert-butylN-(2-{[(3-{5-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohex-1-en-1-yl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate

A suspension oftert-butyl({[6,6-dimethyl-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl]oxy})dimethylsilane(0.15 g, 0.41 mmol), tert-butylN-[2-({[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl}(methyl)amino)ethyl]-N-methylcarbamate(0.13 g, 0.27 mmol) and KOAc (0.11 g, 0.82 mmol) in 1,4-dioxane (10 ml)and water (1 ml) was degassed with a nitrogen sparge for 10 min whilststirring. Pd(dppf)Cl₂.DCM (0.02 g, 0.03 mmol) was added to the reactionmixture which was then stirred at 100° C. for 18 hr in a sealed tube.The reaction was then allowed to cool to RT and evaporated in vacuo. Theresidue was absorbed onto silica gel (˜5 ml). Purification by silica gelcolumn chromatography, on a Biotage Isolera system, using a 25 g KP-SilSNAP cartridge, eluting with EtOAc:heptanes (+1% TEA, 1:9-1), gave thedesired product as a yellow oil (120 mg, 75%): MS (ESI+) forC₃₂H₅₈N₄O₄Si m/z 591.25 (M+H)⁺; HPLC purity 100% (ret. time, 1.39 min);¹H-NMR (500 MHz, Chloroform-d) δ 7.68-7.40 (m, 1H), 6.06-5.73 (m, 1H),5.37-5.27 (m, 1H), 4.09-4.02 (m, 1H), 3.72-3.64 (m, 1H), 3.61 (dd,J=8.0, 5.4 Hz, 1H), 3.48-3.31 (m, 2H), 3.27 (s, 1H), 3.18-3.05 (m, 1H),2.94-2.75 (m, 3H), 2.71-2.61 (m, 1H), 2.54-2.40 (m, 2H), 2.40-2.30 (m,1H), 2.27-2.16 (m, 2H), 2.13-1.95 (m, 5H), 1.66 (d, J=8.9 Hz, 2H),1.49-1.39 (m, 9H), 0.96-0.79 (m, 15H), 0.13-−0.01 (m, 6H).

Step 7: tert-butylN-(2-{[(3-{3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohexyl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate

A mixture of tert-butylN-(2-{[(3-{3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohex-1-en-1-yl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate(120 mg, 0.20 mmol) and 10% Pd—C (0.01 g) in EtOH (10 ml) was stirredunder an atmosphere of hydrogen for 18 hrs. This was filtered andrecharged with another portion of 10% Pd—C and hydrogen and stirredovernight. The reaction mixture was filtered and evaporated in vacuo.Purification by silica gel column chromatography, on a Biotage Isolerasystem, using a 10 g KP-Sil SNAP cartridge, eluting with EtOAc:heptanes(1:9-1), gave the desired product as a colourless glass (49 mg, 40%): MS(ESI+) for C₃₂H₆₀N₄O₄Si m/z 593.25 (M+H)+; HPLC purity 99% (ret. time,1.41 min); ¹H-NMR (500 MHz, Chloroform-d) δ 7.44 (d, J=25.2 Hz, 1H),5.34-5.23 (m, 1H), 4.09-4.00 (m, 1H), 3.74-3.62 (m, 1H), 3.53-3.22 (m,5H), 2.91-2.78 (m, 3H), 2.74-2.63 (m, 1H), 2.45 (s, 2H), 2.19 (d, J=19.7Hz, 3H), 2.07-1.96 (m, 3H), 1.76 (tt, J=13.0, 7.9 Hz, 3H), 1.70-1.48 (m,5H), 1.48-1.39 (m, 10H), 1.34-1.16 (m, 3H), 0.95-0.90 (m, 7H), 0.86 (s,9H).

Step 8:2,2-dimethyl-5-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]cyclohexan-1-ol(Compound 185)

6 N HCl (2 ml) was added to a solution of tert-butylN-(2-{[(3-{3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohexyl}-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino}ethyl)-N-methylcarbamate(49 mg, 0.08 mmol) in 1,4-dioxane (2 ml) at 0° C. and stirred whilstallowing to warm to RT. The reaction was stirred overnight thenevaporated to dryness, then evaporated from MeOH (2×10 ml). The productwas dissolved in MeOH (5 ml) and eluted onto a 2 g Isolute SCX-2cartridge. MeOH (2×10 ml) was eluted then the product was released with7 N NH₃ in MeOH (2×10 ml). This was evaporated to dryness to give ˜20 mgof the product. Purification by silica gel column chromatography, on aBiotage Isolera system, using a 10 g KP-Sil SNAP cartridge, eluting withDCM:MeOH (1:9) and then 7 N NH₃ in MeOH:DCM (1:99-1:9) gave the desiredproduct as a colourless glass (16 mg, 66%): ¹H-NMR (500 MHz,Methanol-d4) δ 7.44 (s, 1H), 3.43 (d, J=8.9 Hz, 2H), 3.40-3.35 (m, 1H),2.84 (ddt, J=12.5, 7.9, 4.0 Hz, 1H), 2.76-2.64 (m, 2H), 2.52 (td, J=6.5,2.5 Hz, 2H), 2.38 (d, J=6.3 Hz, 3H), 2.20 (d, J=3.4 Hz, 3H), 1.76 (ddd,J=24.8, 12.4, 4.2 Hz, 3H), 1.63-1.50 (m, 2H), 1.41-1.26 (m, 2H), 1.04(d, J=7.1 Hz, 3H), 0.98 (d, J=4.2 Hz, 3H); MS (ESI+) for C₁₋₆H₃₀N₄O m/z295.05 (M+H)⁺.

Compound 200 Methyl[2-(methylamino)ethyl]([4-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-3-yl]methyl)aminetrifluoroacetic acid salt

Step 1:(R/S)-4-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-3-carbaldehyde

Into a 50-mL round-bottom flask purged and maintained with an atmosphereof nitrogen was added2-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(500 mg, 1.71 mmol, 1.00 equiv),4-iodo-1-(oxan-2-yl)-1H-pyrazole-3-carbaldehyde (523 mg, 1.71 mmol, 1.00equiv), Pd(dppf)Cl₂ (124 mg, 0.17 mmol, 0.10 equiv), K₃PO₄ (1.087 g,5.12 mmol, 2.99 equiv), ethylene glycol dimethyl ether (10 mL) and water(1 mL). The resulting solution was stirred overnight at 75° C. in an oilbath. The resulting mixture was concentrated under vacuum. The residuewas purified by flash chromatography on silica gel using ethylacetate/petroleum ether (1:4) as eluent to afford 270 mg (46%) of (R/S)4-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-3-carbaldehydeas a yellow oil.

Step 2: (R/S) tert-butylN-(2-[[(4-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-3-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

Into a 50-mL round-bottom flask was placed4-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-3-carbaldehyde(270 mg, 0.78 mmol, 1.00 equiv), tert-butylN-methyl-N-[2-(methylamino)ethyl]carbamate (220 mg, 1.17 mmol, 1.49equiv) and dichlorethane (10 mL). NaBH(OAc)₃ (496 mg, 2.34 mmol, 2.98equiv) was added portionwise and the resultant mixture stirred overnightat room temperature. The reaction was quenched with 15 mL of water andextracted with 3×15 mL of dichloromethane. The combined organic layerswere washed with 20 mL of brine and dried over anhydrous sodium sulfateand then concentrated under vacuum. The residue was purified by flashchromatography on silica gel using dichloromethane/methanol (20:1) toafford 300 mg (74%) of (R/S) tert-butylN-(2-[[(4-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-3-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas a colorless oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.43 (s, 1H), 5.78-5.50(m, 1H), 4.79-4.58 (m, 1H), 4.15-3.77 (m, 2H), 3.77-3.40 (m, 4H),3.40-3.05 (m, 2H), 3.00-2.68 (m, 4H), 2.68-1.99 (m, 10H), 1.99-1.82 (m,2H), 1.82-1.50 (m, 6H), 1.44 (s, 9H), 1.13 (s, 6H) ppm.

Step 3: (R/S) tert-butylN-(2-[[(4-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-3-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

Into a 100-mL round-bottom flask was placed tert-butylN-(2-[[(4-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-3-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(300 mg, 0.58 mmol, 1.00 equiv), 10% Pd(OH)₂/C (300 mg) andtetrahydrofuran (30 mL). The resulting reaction mixture was stirred for1 h at room temperature under 3 atmospheres of hydrogen. The reactionvessel was purged with an inert gas and the mixture filtered under ablanket of inert gas. The filtrate was concentrated under vacuum toprovide 300 mg (100%) of (R/S) tert-butylN-(2-[[(4-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-3-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas a colorless oil. ¹H-NMR (300 MHz, CDCl₃): δ 7.45 (s, 1H), 5.74-5.53(m, 1H), 4.10-3.98 (m, 1H), 3.40-3.25 (m, 3H), 2.79 (s, 3H), 2.60-2.30(m, 5H), 2.30-2.15 (m, 3H), 1.99-1.75 (m, 8H), 1.75-1.50 (m, 10H), 1.42(s, 9H), 1.10 (s, 6H).

Step 4:Methyl[2-(methylamino)ethyl]([4-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-3-yl]methyl)aminetrifluoroacetic acid salt (Compound 200)

Into a 50-mL round-bottom flask was placed (R/S) tert-butylN-(2-[[(4-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-3-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(300 mg, 0.58 mmol, 1.00 equiv), trifluoroacetic acid (5 mL) anddichloromethane (5 mL). The resulting solution was stirred for 30 min atroom temperature and then concentrated under vacuum. The crude product(307 mg) was purified by Prep-HPLC with the following conditions(Prep-HPLC-005(Waters): Column, Atlantis Prep OBD T3 Column, 19×150 mm,5 m; mobile phase, water with 0.05% trifluoroacetic acid and CH₃CN (upto 3.0% in 10 min, up to 100% in 1 min, hold at 100% for 1 min);Detector, UV 220 nm. This resulted in 154.5 mg (50%) of (R/S)methyl[2-(methylamino)ethyl]([4-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-3-yl]methyl)aminetrifluoroacetic acid salt as a colorless solid. ¹H-NMR (300 MHz, D₂O) δ:7.71 (s, 1H), 4.49 (s, 2H), 3.70-3.50 (m, 6H), 2.94 (s, 3H), 2.80 (s,3H), 2.62-2.49 (m, 1H), 2.05-1.87 (m, 2H), 1.79-1.52 (m, 8H), 1.10 (s,6H) ppm.

Compound 205Methyl([1-methyl-3-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)[2-(methylamino)ethyl]aminehydrochloride

Step 1:3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-methyl-1H-pyrazole-4-carbaldehyde

To a stirred mixture of3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1H-pyrazole-4-carbaldehyde(2 g, 7.68 mmol, 1.00 equiv) and potassium carbonate (3.18 g, 23.01mmol, 2.99 equiv) in CH₃CN (40 mL) at 0° C. was added CH₃I (5.46 g,38.47 mmol, 5.01 equiv) dropwise. The resulting mixture was stirredovernight at room temperature then filtered and concentrated undervacuum. The resultant residue was dissolved in H₂O (30 mL) and extractedwith 3×20 mL of dichloromethane. The combined organic layers were washedwith 3×20 mL of brine then dried over anhydrous sodium sulfate. Theresidue was partially purified by flash chromatography on silica gelcolumn using ethyl acetate/petroleum ether (gradient: 1:9-1:2) aseluent. The resultant material was re-purified by Prep-HPLC with thefollowing conditions: Column: XBridge C18, 19×150 mm, 5 μm; Mobile PhaseA: Water/0.05% NH₄HCO₃, Mobile Phase B: ACN; Flow rate: 30 mL/min;Gradient: 20% B to 85% B in 10 min; 254 nm. This resulted in 1.04 g(49%) of3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-methyl-1H-pyrazole-4-carbaldehydeas a white solid. ¹H-NMR (400 MHz, CDCl₃) δ: 9.88 (s, 1H), 7.86 (s, 1H),6.18-6.16 (m, 1H), 3.90 (s, 3H), 3.59-3.54 (m, 3H), 2.78-2.70 (m, 1H),2.59-2.35 (m, 3H), 2.63-2.56 (m, 1H), 1.96-1.90 (m, 1H), 1.75-1.61 (m,2H), 1.13 (s, 6H) ppm. And another isomer, 0.45 g (21.2%) of3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-2-methyl-pyrazole-4-carbaldehyde.¹H-NMR (400 MHz, CDCl₃) δ: 9.70 (s, 1H), 7.91 (s, 1H), 5.84-5.82 (m,1H), 3.80 (s, 3H), 3.60-3.54 (m, 3H), 2.57-2.26 (m, 4H), 2.01-1.94 (m,1H), 1.85-1.80 (m, 1H), 1.78-1.60 (m, 1H), 1.15 (s, 6H) ppm.

Step 2: tert-butylN-(2-[[(3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-methyl-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

To a solution of3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-methyl-1H-pyrazole-4-carbaldehyde(1.04 g, 3.79 mmol, 1.00 equiv) and tert-butylN-methyl-N-[2-(methylamino)ethyl]carbamate (1.07 g, 5.68 mmol, 1.50equiv) in dichloroethane (20 mL) was added NaBH(OAc)₃ (2.41 g)portionwise. The resulting mixture was stirred overnight at 50° C. in anoil bath then cooled to room temperature and quenched by the addition of5 mL of NH₄Cl (sat. aq.). The resulting mixture was dissolved indichloromethane (40 mL) and washed with 3×20 mL of brine then dried overanhydrous sodium sulfate. The residue was purified by flashchromatography on silica gel column using dichloromethane/methanol(20:1) as eluent to afford 1.50 g (89%) of tert-butylN-(2-[[(3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-methyl-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas a yellow oil. ¹H-NMR (300 MHz, D₂O) δ: 8.12 (s, 1H), 5.72-5.70 (m,1H), 4.25-3.94 (m, 4H), 3.75-3.34 (m, 5H), 3.16-2.88 (m, 4H), 2.51-2.04(m, 7H), 1.93-1.62 (m, 5H), 1.45 (s, 9H), 1.13 (s, 6H).

Step 3:N-methyl-N-[2-[methyl([1-methyl-3-[(5s,8s)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amino]ethyl]carbamate

Into a 250-mL round-bottom flask was placed tert-butylN-(2-[[(3-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-methyl-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(1.50 g, 3.36 mmol, 1.00 equiv), tetrahydrofuran (30 mL), and 10%Pd(OH)₂/C (1.50 g). The resulting mixture was stirred overnight at roomtemperature under 3 atmospheres of hydrogen. The resulting mixture wasfiltered then concentrated under vacuum. The residue was partiallypurified by flash chromatography on silica gel column usingdichloromethane/methanol (20:1) as eluent. The partially purifiedmaterial was then repurified under the following conditions: Column:XBridge C18, 19×150 mm, 5 μm; Mobile Phase A: Water/0.05% NH₄HCO₃,Mobile Phase B: ACN; Flow rate: 30 mL/min; Gradient: 20% B to 85% B in10 min; 254 nm. This resulted in 1.0 g (66%) of tert-butylN-methyl-N-[2-[methyl([1-methyl-3-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amino]ethyl]carbamateas a yellow oil. ¹H-NMR (400 MHz, CD₃Cl) δ: 7.14 (s, 1H), 3.80-3.78 (m,4H), 3.51 (s, 3H), 3.36-3.28 (m, 3H), 2.85 (s, 3H), 2.58-2.46 (m, 2H),2.28-2.22 (m, 3H), 2.01-1.84 (m, 4H), 1.68-1.53 (m, 4H), 1.55 (s, 2H),1.43 (s, 9H), 1.13 (s, 6H).

Step 4:Methyl([1-methyl-3-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)[2-(methylamino)ethyl]aminehydrochloride (Compound 205)

Into a 25-mL round-bottom flask was placed tert-butylN-methyl-N-[2-[methyl([1-methyl-3-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amino]ethyl]carbamate(230 mg, 0.51 mmol, 1.00 equiv) and dichloromethane (10 mL). Hydrogenchloride gas was bubbled through the reaction mixture. The reactionmixture was then stirred for 0.5 h at room temperature then extractedwith 3×10 mL of water and the aqueous layers combined and concentratedunder vacuum. This resulted in 131 mg (61%) ofmethyl([1-methyl-3-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)[2-(methylamino)ethyl]aminehydrochloride as a colorless solid. ¹H-NMR (400 MHz, D₂O): 7.66 (s, 1H),4.25 (s, 2H), 3.74 (s, 3H), 3.50-3.40 (m, 6H), 2.74 (s, 3H), 2.70 (s,3H), 2.64-2.54 (m, 1H), 2.60 (s, 1H), 1.89 (d, 2H, J=16 Hz), 1.70-1.42(m, 8H), 0.99 (s, 6H). LCMS (method A5, ESI): RT=1.33 min, m/z=349.2[M+H]⁺.

Compound 206Methyl([1-methyl-3-[(5R,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)[2-(methylamino)ethyl]aminehydrochloride

Step 1:Methyl([1-methyl-3-[(5R,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)[2-(methylamino)ethyl]aminehydrochloride (Compound 206)

Into a 25-mL round-bottom flask was placed tert-butylN-methyl-N-[2-[methyl([1-methyl-3-[(5R,8R)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amino]ethyl]carbamate(80 mg, 178.32 mmol, 1.00 equiv) and dichloromethane (10 mL). Hydrogenchloride (gas) was bubbled through the reaction mixture. The reactionmixture was then stirred overnight at room temperature and thenextracted with 3×10 mL of water and the aqueous layers combined andconcentrated under vacuum. This resulted in 23.1 mg ofmethyl([1-methyl-3-[(5R,8R)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)[2-(methylamino)ethyl]aminehydrochloride as a colorless solid. ¹H-NMR (400 MHz, D₂O) δ: 7.66 (s,1H), 4.25 (s, 2H), 3.74 (s, 3H), 3.50-3.40 (m, 6H), 2.74 (s, 3H), 2.70(s, 3H), 2.67-2.55 (m, 1H), 1.87-1.68 (m, 6H), 1.52-1.42 (m, 4H), 1.02(s, 6H). LCMS (method A5, ESI): RT=1.32 min, m/z=349.15 [M+H]⁺.

Compound 245 Methyl([2-[methyl([1-methyl-5-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amino]ethyl])aminetrifluoroacetic acid salt

Step 1: 3-Iodo-1H-pyrazole-4-carbaldehyde

To stirred solution of 3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde(3 g, 9.80 mmol, 1.00 equiv) in dichloromethane (10 mL) was addedtrifluoroacetic acid (10 mL). The resulting solution was stirred for 3 hat room temperature then concentrated under vacuum and the resultingresidue was treated with sufficient sodium carbonate (sat. aq.) solutionto afford a mixture of pH 8. The resulting solution was extracted with50 mL of dichloromethane and the organic layer washed with brine (3×50mL) and dried over anhydrous sodium sulfate and concentrated undervacuum. The residue was purified by flash chromatography on silica gelusing dichloromethane/petroleum ether (1:3) as eluent to afford 1.4 g(64%) of 3-iodo-1H-pyrazole-4-carbaldehyde as a white solid. ¹H-NMR (300MHz, CDCl₃): δ 9.89 (s, 1H), 8.04 (s, 1H) ppm.

Step 2: 5-iodo-1-methyl-1H-pyrazole-4-carbaldehyde

To a stirred solution of 3-iodo-1H-pyrazole-4-carbaldehyde (1.4 g, 6.31mmol, 1.00 equiv) in CH₃CN (20 mL) at 0° C. was added potassiumcarbonate (2.5 g, 18.09 mmol, 2.87 equiv) followed by dropwise additionof CH₃I (980 mg, 6.90 mmol, 1.09 equiv). The resulting mixture wasstirred for 3 h at room temperature, then filtered and concentratedunder vacuum. The residue was purified by flash chromatography on silicagel using ethyl acetate/petroleum ether (1:10) as eluent to afford 400mg (27%) of 5-iodo-1-methyl-1H-pyrazole-4-carbaldehyde as a white solid.¹H-NMR (300 MHz, CDCl₃): δ 9.61 (s, 1H), 8.02 (s, 1H), 3.92 (s, 3H) ppm.

Step 3:5-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-methyl-1H-pyrazole-4-carbaldehyde

Into a 50-mL 3-necked round-bottom flask purged and maintained with aninert atmosphere of nitrogen was placed2-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(686 mg, 2.35 mmol, 1.00 equiv), 1,4-dioxane (20 mL),5-iodo-1-methyl-1H-pyrazole-4-carbaldehyde (370 mg, 1.57 mmol, 0.67equiv), Pd(dppf)Cl₂ (115 mg, 0.16 mmol, 0.07 equiv), water (2 mL) andpotassium carbonate (650 mg, 4.70 mmol, 2.00 equiv). The resultingmixture was stirred overnight at 80° C. then concentrated under vacuum.The residue was purified by flash chromatography on silica gel columnusing ethyl acetate/petroleum ether (1:7) as eluent to afford 340 mg(53%) of5-[3,3-dimethyl-1-oxaspiro[4.5]dec-7-en-8-yl]-1-methyl-1H-pyrazole-4-carbaldehydeas a light yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ 9.70 (s, 1H), 7.92 (s,1H), 5.84-5.83 (m, 1H), 3.80 (s, 3H), 3.60 (s, 2H), 2.57-2.26 (m, 4H),2.04-1.68 (m, 4H), 1.21-1.10 (m, 6H) ppm.

Step 4:(5-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-methyl-1H-pyrazol-4-yl)methanol

Into a 50-mL round-bottom flask was placed5-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-methyl-1H-pyrazole-4-carbaldehyde(340 mg, 1.23 mmol, 1.00 equiv), tetrahydrofuran (20 mL) and 10%Pd(OH)₂/C (680 mg). The resulting mixture was stirred overnight at roomtemperature under 3 atmospheres of hydrogen then filtered andconcentrated under vacuum to afford 340 mg of(5-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-methyl-1H-pyrazol-4-yl)methanolas light yellow oil.

Step 5: tert-butylN-(2-[[(5-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-methyl-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

To a solution of(5-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-methyl-1H-pyrazol-4-yl)methanol(340 mg, 1.22 mmol, 1.00 equiv) and triethylamine (371 mg, 3.67 mmol,3.00 equiv) in dichloromethane (8 mL) at 0° C. was added methanesulfonylchloride (167.3 mg) dropwise with stirring. The resulting solution wasstirred for 30 min at room temperature then treated with tert-butylN-methyl-N-[2-(methylamino)ethyl]carbamate (276 mg, 1.47 mmol, 1.20equiv). The resulting solution was stirred for 2 h at room temperaturethen quenched by the addition of 20 mL of water. The resulting mixturewas extracted with 3×50 mL of dichloromethane and the organic layerscombined and dried over anhydrous sodium sulfate and concentrated undervacuum to afford 128 mg (23%) of tert-butylN-(2-[[(5-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-methyl-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas a light yellow oil. ¹H-NMR (300 MHz, CD₃OD): δ 7.30 (s, 1H), 3.83 (s,3H), 3.53 (s, 2H), 3.43-3.31 (m, 4H), 2.91-2.83 (m, 4H), 2.53-2.50 (m,2H), 2.25-2.07 (m, 5H), 1.97-1.93 (m, 2H), 1.63-1.49 (m, 15H), 1.18-1.10(m, 6H) ppm.

Step 6: Methyl([2-[methyl([1-methyl-5-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amino]ethyl])aminetrifluoroacetic acid salt (Compound 245)

Into a 8-mL sealed tube was placed tert-butylN-(2-[[(5-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1-methyl-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(37 mg, 0.08 mmol, 1.00 equiv), dichloromethane (1 mL) and CF₃COOH (1mL). The resulting solution was stirred for 30 min at room temperaturethen concentrated under vacuum. The resultant residue was purified byreverse phase HPLC using the following conditions: Column: Sunfire C18,19×150 mm, 5 μm; Mobile Phase A: Water/0.05% TFA, Mobile Phase B: ACN;Flow rate: 30 mL/min; Gradient: 5% B to 55% B in 10 min; 254 nm. Thisresulted in 18.9 mg (40%) of methyl([2-[methyl([1-methyl-5-[(5S,8S)-3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-4-yl]methyl)amino]ethyl])aminetrifluoroacetic acid salt as a colorless oil. ¹H-NMR (300 MHz, D₂O): δ7.53 (s, 1H), 4.33 (s, 2H), 3.81 (s, 3H), 3.52-3.40 (m, 6H), 2.91-2.69(m, 7H), 2.00-1.72 (m, 4H), 1.69-1.45 (m, 6H), 1.00 (s, 6H). LCMS(method W, ESI): RT=1.37 min, m/z=349.1 [M+H].

Compound 2174-[4-([methyl[2-(methylamino)ethyl]amino]methyl)-1H-pyrazol-3-yl]cyclohexan-1-oltrifluoroacetate

Step 1: 1,4-dioxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate

To a solution of 1,4-dioxaspiro[4.5]decan-8-one (80 g, 512.23 mmol, 1.00equiv) in THF (500 mL) at −78° C. was added LiHMDS (615 mL of a 1 Msolution in THF) dropwise over approximately 25 min then stirred for 2 hat −40° C. The reaction mixture was then cooled to −78° C. and treatedwith1,1,1-trifluoro-N-phenyl-N-(trifluoromethane)sulfonylmethanesulfonamide(220 g, 615.82 mmol, 1.20 equiv) dropwise. The resulting solution wasallowed to warm to room temperature and stirred overnight then quenchedby the addition of 100 mL of NH₄Cl (sat. aq.). The resulting mixture wasextracted with 500 mL of ethyl acetate and the organic extract washedwith 3×500 mL of brine and dried over anhydrous sodium sulfate. Thecrude product was purified by flash chromatography on silica gel columnusing ethyl acetate/petroleum ether (gradient: 1% to 3% EA) as eluent toafford 166 g of 1,4-dioxaspiro[4.5]dec-7-en-8-yltrifluoromethanesulfonate as a yellow oil. ¹H-NMR (300 MHz, CDCl₃):5.68-5.64 (m, 1H), 3.99 (s, 4H), 2.56-2.51 (m, 2H), 2.42-2.41 (m, 2H),1.90 (t, J=6.6 Hz, 2H) ppm.

Step 2:2-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

Into a 3-L 4-necked round-bottom flask was placed1,4-dioxaspiro[4.5]dec-7-en-8-yl trifluoromethanesulfonate (80 g, 277.55mmol, 1.00 equiv), B₂Pin₂ (85 g, 334.65 mmol, 1.21 equiv), Pd(dppf)Cl₂(20 g, 27.33 mmol, 0.10 equiv), KOAc (82 g, 835.54 mmol, 3.01 equiv) and1,4-dioxane (800 mL). The resulting solution was stirred overnight at80° C. using an oil bath then cooled to room temperature andconcentrated under vacuum. The residue was extracted with 1 L of ethylacetate and the organic layer washed with 3×1 L of brine and dried overanhydrous sodium sulfate and concentrated under vacuum. The residue waspurified by flash chromatography on silica gel column using ethylacetate/petroleum ether (gradient: 5% to 10% ethyl acetate) to afford 36g (49%) of2-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolaneas a yellow solid. ¹H-NMR (300 MHz, CDCl₃): 6.46-6.47 (m, 1H), 3.98 (s,4H), 2.39-2.35 (m, 4H), 1.73 (t, J=4.8 Hz, 2H), 1.26 (s, 12H) ppm.

Step 3:3-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde

Into a 2-L 4-necked round-bottom flask was placed2-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(44 g, 165.33 mmol, 1.00 equiv) and3-iodo-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde (45.5 g, 148.64 mmol,0.90 equiv). This was followed by the addition of Pd(dppf)Cl₂ (12 g,16.40 mmol, 0.10 equiv). To this was added K₃PO₄ (105 g, 494.66 mmol,2.99 equiv), ethylene glycol dimethyl ether (500 mL) and water (50 mL).The resulting mixture was stirred overnight at 75° C. in an oil baththen cooled to room temperature and concentrated under vacuum. Theresidue was extracted with 500 mL of ethyl acetate and the organic layerwashed with 3×500 mL of brine and dried over anhydrous sodium sulfateand concentrated under vacuum. The residue was purified by flashchromatography on silica gel column using ethyl acetate/petroleum ether(gradient: 20% to 30% EA) as eluent to afford 35.5 g (67%) of (R/S)3-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehydeas a light yellow solid. ¹H-NMR (300 MHz, CDCl₃): 9.91 (s, 1H), 8.26 (s,1H), 6.29-6.26 (m, 1H), 5.38-5.34 (m, 1H), 4.09-4.01 (m, 5H), 3.74-3.63(m, 1H), 2.79-2.74 (m, 2H), 2.50-2.49 (d, J=3.6 Hz, 2H), 2.12-1.89 (m,5H), 1.72-1.60 (m, 3H) ppm.

Step 4: (R/S) tert-butylN-(2-[[(3-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

To a stirred solution of (R/S)3-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazole-4-carbaldehyde(25 g, 78.53 mmol, 1.00 equiv) and tert-butylN-methyl-N-[2-(methylamino)ethyl]carbamate (17.7 g, 94.02 mmol, 1.20equiv) in dichloroethane (250 mL) at 0° C. was added NaBH(OAc)₃ (50 g,235.91 mmol, 3.00 equiv) portionwise. The resulting mixture was allowedto warm to room temperature, stirred overnight and then quenched by theaddition of 50 mL of NH₄Cl (sat. aq.). The resulting mixture wasextracted with 500 mL of CH₂Cl₂ and the organic phase washed with 3×500mL of brine and dried over anhydrous sodium sulfate and concentratedunder vacuum. The residue was purified by flash chromatography on silicagel column using ethyl acetate/petroleum ether (1:2) as eluent to afford30.3 g (79%) of (R/S) tert-butylN-(2-[[(3-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas a yellow oil. ¹H-NMR (300 MHz, CDCl₃): 7.50 (s, 1H), 6.26 (br, 1H),5.38-5.34 (m, 1H), 4.09-3.99 (m, 5H), 3.74-3.69 (m, 1H), 3.42 (br, 2H),2.86 (s, 3H), 2.84-2.07 (m, 8H), 2.04 (s, 3H), 1.85 (t, J=6.6 Hz, 2H),1.68-1.52 (m, 6H), 1.58 (s, 9H) ppm.

Step 5: (R/S) tert-butylN-(2-[[(3-[1,4-dioxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

Into a 1-L round-bottom flask was placed (R/S) tert-butylN-(2-[[(3-[1,4-dioxaspiro[4.5]dec-7-en-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(15 g, 30.57 mmol, 1.00 equiv), THF (500 mL) and 10% Pd(OH)2/C (9 g).The resulting mixture was stirred for 2 h at room temperature under 3atmospheres of hydrogen. The resulting mixture was filtered andconcentrated under vacuum to afford 11.5 g (76%) of (R/S) tert-butylN-(2-[[(3-[1,4-dioxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas yellow oil. ¹H-NMR (300 MHz, CDCl3): 7.41 (s, 1H), 5.30-5.25 (m, 1H),4.11-3.95 (m, 5H), 3.70-3.62 (m, 1H), 3.36 (br, 4H), 2.83 (s, 3H),2.74-2.66 (m, 1H), 2.47 (s, 3H), 2.04 (s, 3H), 2.04-1.82 (m, 10H),1.68-1.52 (m, 6H), 1.48 (s, 9H) ppm.

Step 6: (R/S) tert-butylN-methyl-N-[2-[methyl([[1-(oxan-2-yl)-3-(4-oxocyclohexyl)-1H-pyrazol-4-yl]methyl])amino]ethyl]carbamate

Into a 250-mL round-bottom flask was placed tert-butylN-(2-[[(3-[1,4-dioxaspiro[4.5]decan-8-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(13.5 g, 27.40 mmol, 1.00 equiv), dichloromethane (130 mL) andFeCl₃-6H₂O (26 g, 96.30 mmol, 3.51 equiv). The resulting solution wasstirred for 2 h at room temperature then extracted with dichloromethane(200 mL). The organic phase was washed sequentially with 3×100 mL ofbrine, 3×100 mL sodium bicarbonate (sat. aq.) and then again with 3×100mL of brine. The mixture was dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was purified by flashchromatography on silica gel column using methanol/dichloromethane(gradient: 1% to 5% MeOH) to afford 7.5 g (61%) of tert-butylN-methyl-N-[2-[methyl([[1-(oxan-2-yl)-3-(4-oxocyclohexyl)-1H-pyrazol-4-yl]methyl])amino]ethyl]carbamateas a yellow oil.

Step 7:(R/S)N-[2-([[3-(4-hydroxycyclohexyl)-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl](methyl)amino)ethyl]-N-methylcarbamate

To a stirred solution of (R/S) tert-butylN-methyl-N-[2-[methyl([[1-(oxan-2-yl)-3-(4-oxocyclohexyl)-1H-pyrazol-4-yl]methyl])amino]ethyl]carbamate(500 mg, 1.11 mmol) in methanol (5 mL) at 0° C. was added NaBH₄ (85 mg,2.24 mmol) portionwise. The resulting mixture was stirred for 1 h atroom temperature then quenched by the addition of 5 mL of NH₄Cl (sat.aq.). The resulting mixture was concentrated under vacuum to afford 380mg (76%) of (R/S) tert-butylN-[2-([[3-(4-hydroxycyclohexyl)-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl](methyl)amino)ethyl]-N-methylcarbamateas a light yellow oil. ¹H-NMR (400 MHz, CDCl₃): δ 7.41 (s, 1H), 5.28 (t,J=6.0 Hz, 1H), 4.05 (d, J=8.0 Hz, 1H), 3.74-3.62 (m, 2H), 3.41-3.25 (m,3H), 2.84 (s, 3H), 2.68-2.56 (m, 2H), 2.20 (s, 3H), 2.09-1.82 (m, 6H),1.74-1.52 (m, 7H), 1.51-1.29 (m, 13H) ppm.

Step 8: tert-butyl2-(((3-((1S,4S)-4-hydroxycyclohexyl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl(methyl)carbamate

A solution of (R/S) tert-butylN-[2-([[3-(4-hydroxycyclohexyl)-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl](methyl)amino)ethyl]-N-methylcarbamate(380 mg, 0.84 mmol, 1.00 equiv) in methanol (30 mL) was treated withaqueous hydrochloric acid (12N, 0.06 mL) and stirred overnight at roomtemperature. The pH value of the solution was adjusted to 7-8 withammonia and the mixture dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was purified by reverse phaseHPLC with the following conditions: Column: Sunfire C18, 19×150 mm, 5μm; Mobile Phase A: Water/0.05% ammonium hydroxide Mobile Phase B: ACN;Flow rate: 30 mL/min; Gradient: 5% B to 55% B in 10 min; Detector: 254nm. This resulted in the cis-isomer 30 mg of tert-butyl2-(((3-((1S,4S)-4-hydroxycyclohexyl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl(methyl)carbamate.LCMS (method D, ESI): RT=1.12 min, m/z=367.0 [M+H]⁺. And thetrans-isomer 130 mg of tert-butyl2-(((3-((1R,4R)-4-hydroxycyclohexyl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl(methyl)carbamateas a light yellow oil.

Step 9:4-[4-([methyl[2-(methylamino)ethyl]amino]methyl)-1H-pyrazol-3-yl]cyclohexan-1-oltrifluoroacetate (Compound 217)

A solution of tert-butyl2-(((3-((1S,4S)-4-hydroxycyclohexyl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl(methyl)carbamate(30 mg, 0.08 mmol, 1.00 equiv) in dichloromethane (2 mL) was treatedwith trifluoroacetic acid (2 mL) and stirred for 5 min at roomtemperature. The resulting mixture was concentrated under vacuum toafford 28.6 mg (71%) of4-[4-([methyl[2-(methylamino)ethyl]amino]methyl)-1H-pyrazol-3-yl]cyclohexan-1-oltrifluoroacetate salts as a light yellow oil. ¹H-NMR (300 MHz, D₂O): δ7.70 (s, 1H), 4.30 (s, 2H), 4.70-4.00 (m, 1H), 3.51-3.40 (m, 4H),2.82-2.66 (m, 7H), 1.86-1.52 (m, 8H) ppm. LCMS (method A6, ESI): RT=2.78min, m/z=267.05 [M+H]⁺.

Compound 2274-[4-([methyl[2-(methylamino)ethyl]amino]methyl)-1H-pyrazol-3-yl]cyclohexan-1-oltrifluoroacetate

Step 1:4-[4-(I[methyl[2-(methylamino)ethyl]amino]methyl)-1H-pyrazol-3-yl]cyclohexan-1-oltrifluoroacetate (Compound 227)

A solution of tert-butyl2-(((3-((1R,4R)-4-hydroxycyclohexyl)-1H-pyrazol-4-yl)methyl)(methyl)amino)ethyl(methyl)carbamate(130 mg, 0.35 mmol, 1.00 equiv)) in dichloromethane (2 mL) was treatedwith trifluoroacetic acid (2 mL) and stirred for 5 min at roomtemperature. The resulting mixture was concentrated under vacuum toafford 120.9 mg (69%) of4-[4-([methyl[2-(methylamino)ethyl]amino]methyl)-1H-pyrazol-3-yl]cyclohexan-1-oltrifluoroacetate as a light yellow oil. ¹H-NMR (300 MHz, D₂O): δ 7.70(s, 1H), 4.30 (s, 2H), 3.71-3.60 (m, 1H), 3.51-3.41 (m, 4H), 2.78-2.60(m, 7H), 2.01-1.91 (m, 2H), 1.87-1.75 (m, 2H), 1.61-1.45 (m, 2H),1.40-1.22 (m, 2H) ppm. LCMS (method A6, ESI): RT=2.83 min, m/z=267.1[M+H]⁺.

Compound 263(1S)-2,2-dimethyl-5-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]-N-(3-methylbutyl)cyclohexane-1-carboxamide

Step 1: (2S)-2-(methoxymethyl)-N-methylidenepyrrolidin-1-amine

To an ice cold solution of (2S)-2-(methoxymethyl)pyrrolidin-1-amine (5.0g, 38.41 mmol) in DCM (75 mL) was added paraformaldehyde (1.38 g, 46.09mmol). The mixture was left to stir at RT over the weekend. Water (25mL) was added, and the phases were separated. The aqueous phase wasextracted with DCM (3×30 mL). The combined organics were washed withwater (20 ml), brine (20 ml), dried over Na₂SO₄, filtered and evaporatedto dryness. Purification by silica gel column chromatography, on aBiotage Isolera system, using a 100 g HP-Sil SNAP cartridge, elutingwith EtOAc:heptane (1:99-4:6), gave the desired product as a colourlessoil (4.19 g, 76%): MS (ESI⁺) for C₇H₁₄N₂O m/z 143.0 (M+H)⁺.; HPLC purity100% (ret. time, 0.81 min); ¹H-NMR (500 MHz, Chloroform-d) δ 6.13 (d,J=11.6 Hz, 1H), 6.02 (d, J=11.6 Hz, 1H), 3.62-3.51 (m, 2H), 3.49-3.42(m, 1H), 3.38 (s, 3H), 3.33 (ddd, J=9.9, 7.3, 3.4 Hz, 1H), 2.83 (q,J=7.9 Hz, 1H), 2.04-1.87 (m, 3H), 1.86-1.75 (m, 1H).

Step 2:(3S)-3-[(E)-N-[(2S)-2-(methoxymethyl)pyrrolidin-1-yl]carboximidoyl]-4,4-dimethylcyclohexan-1-one

To a cooled (−78° C.) solution of 4,4-dimethylcyclohex-2-en-1-one (4.57g, 36.83 mmol) in dry THF (100 mL) were sequentially addedtert-butyl(dimethyl)silyl trifluoromethanesulfonate (7.45 ml, 32.41mmol) and pre-cooled (−78° C.)(2S)-2-(methoxymethyl)-N-methylidenepyrrolidin-1-amine (4.19 g, 29.47mmol) under a N₂ atmosphere. After 45 min 1MN,N,N-tributylbutan-1-aminium fluoride (44.20 ml, 44.20 mmol) was addedand the mixture was allowed to warm to RT and stirred until LC/MSindicated total consumption of the silyl enol ether (overnight). Thereaction mixture was diluted with t-butylmethylether (100 ml) and washedwith water (2×100 ml). The aqueous was then extracted witht-butylmethylether (100 ml). The combined organic layers were washedwith brine (100 ml), dried over Na₂SO₄, filtered and evaporated in vacuoto give a dark brown oil. Purification by silica gel columnchromatography, on a Biotage Isolera system, using a 340 g KP-Sil SNAPcartridge, eluting with EtOAc:heptanes (1:9-6:4), gave the desiredproduct as a yellow oil (4.6 g, 59%): MS (ESI⁺) for C₁₅H₂₆N₂O₂ m/z266.95 (M+H)⁺.; HPLC purity 100% (ret. time, 1.11 min); ¹H-NMR (500 MHz,Chloroform-d) δ 6.60-6.51 (m, 1H), 3.56-3.49 (m, 1H), 3.47-3.41 (m, 1H),3.40-3.28 (m, 5H), 2.73 (q, J=8.0 Hz, 1H), 2.56-2.25 (m, 5H), 2.01-1.84(m, 3H), 1.83-1.70 (m, 2H), 1.67-1.60 (m, 1H), 1.07 (d, J=11.2 Hz, 6H).

Step 3: 3 (1S)-2,2-dimethyl-5-oxocyclohexane-1-carbaldehyde

A solution of(3S)-3-[(E)-N-[(2S)-2-(methoxymethyl)pyrrolidin-1-yl]carboximidoyl]-4,4-dimethylcyclohexan-1-one(13.7 g, 51.43 mmol) in DCM (250 ml) was cooled to −78° C. and dry ozonewas bubbled through until appearance of a permanent green/blue colour(˜4 h) and then continued for a further 30 min. The reaction mixture wassparged with nitrogen for 20 min. Dimethylsulfide (3.9 ml, 62.13 mmol)was added and the reaction mixture stirred at RT for 30 min beforeevaporating in vacuo. Purification by silica gel column chromatography,on a Biotage Isolera system, using a 340 g KP-Sil SNAP cartridge,eluting with EtOAc:heptanes (1:9-7:3), gave the desired product as ayellow oil (4.02 g, 46%): ¹H-NMR (500 MHz, Chloroform-d) δ 9.85 (d,J=1.5 Hz, 1H), 2.67-2.62 (m, 1H), 2.61-2.54 (m, 1H), 2.48-2.40 (m, 1H),2.39-2.29 (m, 2H), 1.78-1.71 (m, 2H), 1.32 (s, 3H), 1.15 (s, 3H).

Step 4: (1S)-2,2-dimethyl-5-oxocyclohexane-1-carboxylic acid

(1S)-2,2-Dimethyl-5-oxocyclohexane-1-carbaldehyde (4.02 g, 23.46 mmol)in ether (200 ml) was cooled to −30° C. and treated with 2Mtrioxochromium-sulfuric acid (1:1) (58.66 ml, 117.31 mmol Jones'Reagent). After 30 min at −30° C. the mixture was stirred for a further2 hr whilst allowing to warm to RT. The reaction mixture was cooled to0° C. and basified with 1N NaOH (˜650 ml) and washed witht-butylmethylether (˜2×500 ml). The aqueous layer was acidified to acidpH with 2 N H₂SO₄ (˜160 ml) and the product was extracted with EtOAc(3×800 ml). The combined organic layers were dried over Na₂SO₄,filtered, concentrated and co-evaporated with heptane (˜50 ml).Purification by silica gel column chromatography, on a Biotage Isolerasystem, using a 100 g KP-Sil SNAP cartridge, eluting with MeOH:DCM(1:99-1:9), gave the desired product as an off-white solid (3.04 g,76%): ¹H-NMR (500 MHz, Chloroform-d) δ 2.73-2.57 (m, 2H), 2.51-2.40 (m,2H), 2.38-2.31 (m, 1H), 1.95-1.86 (m, 1H), 1.72-1.63 (m, 1H), 1.22 (s,3H), 1.17 (s, 3H).

Step 5: methyl (1S)-2,2-dimethyl-5-oxocyclohexane-1-carboxylate

MeI (1.22 mL, 19.65 mmol) was added to a suspension of(1S)-2,2-dimethyl-5-oxocyclohexane-1-carboxylic acid (3.04 g, 17.86mmol) and K₂CO₃ (2.72 g, 19.65 mmol) in acetone (45 mL) and heated to60° C. for 18 h. This was then allowed to cool to RT, filtered usingadditional DCM 2×20 ml, and evaporated in vacuo. Purification by silicagel column chromatography, on a Biotage Isolera system, using a 100 gKP-Sil SNAP cartridge, eluting with EtOAc:heptanes (1:9-1), gave thedesired product as a colourless oil (2.80 g, 85%): ¹H-NMR (250 MHz,Chloroform-d) δ 3.69 (s, 3H), 2.73-2.54 (m, 2H), 2.53-2.24 (m, 3H),1.95-1.80 (m, 1H), 1.74-1.59 (m, 1H), 1.16 (s, 3H), 1.11 (s, 3H).

Step 6: methyl(1S)-6,6-dimethyl-3-(trifluoromethanesulfonyloxy)cyclohex-3-ene-1-carboxylate

To a cold [0° C.] solution of methyl(1S)-2,2-dimethyl-5-oxocyclohexane-1-carboxylate (1.7 g, 9.27 mmol) in1,2-dichloroethane (50 ml) was added 2,6-di-tert-butylpyridine (2.28 ml,10.15 mmol) followed by slow addition of a solution of Tf₂O (1.65 ml,9.79 mmol). The reaction was allowed to warm to RT overnight. Thesolvent was evaporated and the residue was partitioned between water (50ml) and t-butylmethylether-EtOAc (120 ml, ˜10:1). The organic layer wasseparated, washed with water (25 ml), sat NaHCO₃ (25 ml), brine (25 ml),dried over Na₂SO₄, filtered and concentrated. The residue was absorbedonto silica gel, and purified by silica gel column chromatography,eluting with EtOAc:heptanes (0-1:4) to give the desired product as ayellow oil (2.1 g, 72%): ¹H-NMR (500 MHz, Chloroform-d) δ 5.71 (t, J=3.9Hz, 1H), 2.73-2.62 (m, 1H), 2.60-2.53 (m, 1H), 2.50-2.39 (m, 1H),2.12-2.04 (m, 2H), 1.04 (s, 3H), 1.01 (s, 3H).

Step 7: methyl(1S)-6,6-dimethyl-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate

A suspension of methyl(1S)-6,6-dimethyl-3-(trifluoromethanesulfonyloxy)cyclohex-3-ene-1-carboxylate(2.10 g, 6.64 mmol), KOAc (4.89 g, 49.80 mmol) and4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (2.02 g, 7.97mmol) in 1,4-dioxane (120 ml) was degassed for 10 min under a nitrogensparge at RT. Pd(dppf).Cl₂ (0.04 g, 0.05 mmol) was added to the reactionmixture and stirred at 90° C. for 3 h, then allowed to stir whilstcooling to RT. The reaction mixture was diluted with EtOAc (130 ml) andwashed with water (130 ml). The aqueous layer was extracted with EtOAc(130 ml), the combined organics washed with brine (50 ml), dried overNa₂SO₄, filtered and evaporated in vacuo. Purification by silica gelcolumn chromatography, on a Biotage Isolera system, using a 100 g KP-SilSNAP cartridge, eluting with EtOAc:heptanes (1:99-3:7), gave the desiredproduct as a white solid (1.62 g, 83%): ¹H-NMR (250 MHz, Chloroform-d) δ6.59-6.39 (m, 1H), 3.64 (s, 3H), 2.36 (s, 3H), 2.07-1.90 (m, 2H), 1.25(s, 12H), 0.96 (d, J=4.2 Hz, 6H).

Step 8: methyl(1S)-3-(4-{[(2-{[(tert-butoxy)carbonyl](methyl)amino}ethyl)(methyl)amino]methyl}-1-(oxan-2-yl)-1H-pyrazol-3-yl)-6,6-dimethylcyclohex-3-ene-1-carboxylate

A suspension of methyl(1S)-6,6-dimethyl-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-ene-1-carboxylate(1.62 g, 5.51 mmol), tert-butylN-[2-({[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl}(methyl)amino)ethyl]-N-methylcarbamate(2.63 g, 5.51 mmol), K₂CO₃ (2.30 g, 16.63 mmol) and Pd(dppf)Cl₂.DCM(0.45 g, 0.55 mmol) in 1,4-dioxane (100 ml) and water (10 ml) wasstirred under a N₂ sparge for 10 min at RT. This was then heated to 90°C. and stirred overnight under N₂. The reaction mixture was allowed tocool to RT before evaporating to dryness. MeOH (2×20 ml) was added tothe residue and evaporated to dryness in vacuo. Purification by silicagel column chromatography, on a Biotage Isolera system, using a 100 gKP-Sil SNAP cartridge, eluting with THF:heptanes (1:99-12:88), gave thedesired product as a tan oil (1.89 g, 57%): MS (ESI+) for C₂₈H₄₆N₄O₅ m/z519.10 (M+H)⁺.; HPLC purity 86% (ret. time, 1.14 min); ¹H-NMR (500 MHz,Chloroform-d) δ 7.56-7.38 (m, 1H), 6.12 (s, 1H), 5.34-5.23 (m, 1H), 4.05(d, J=10.0 Hz, 1H), 3.73-3.59 (m, 4H), 3.45-3.22 (m, 4H), 2.82 (s, 3H),2.77-2.63 (m, 2H), 2.57-2.39 (m, 3H), 2.21 (s, 3H), 2.16-1.95 (m, 5H),1.73-1.56 (m, 3H), 1.44 (s, 9H), 1.03 (m, 6H).

Step 9: methyl(1S)-5-(4-{[(2-{[(tert-butoxy)carbonyl](methyl)amino}ethyl)(methyl)amino]methyl}-1-(oxan-2-yl)-1H-pyrazol-3-yl)-2,2-dimethylcyclohexane-1-carboxylate

10% Pd—C (189 mg) was added to a solution of methyl(1S)-3-(4-{[(2-{[(tert-butoxy)carbonyl](methyl)amino}ethyl)(methyl)amino]methyl}-1-(oxan-2-yl)-1H-pyrazol-3-yl)-6,6-dimethylcyclohex-3-ene-1-carboxylate(1.89 mg, 3.13 mmol) in EtOH (30 ml) and stirred under an atmosphere ofhydrogen for 3 h. An additional 189 mg of 10% Pd—C was added and thereaction continued overnight. After stirring overnight an additional 189mg of 10% Pd—C was added and the reaction continued for 3 h. This wasthen filtered and re-treated with 10% Pd—C (190 mg) and hydrogen for 4h. The reaction mixture was filtered and allowed to stand over theweekend. The reaction mixture was then treated with 10% Pd—C (0.5 g) andhydrogen for a further 48 hrs before filtering through Celite andevaporating to dryness. Purification by silica gel columnchromatography, on a Biotage Isolera system, using a 100 g KP-Sil SNAPcartridge, eluting with THF:heptanes (1:9-1), gave the desired productas a colourless oil (1.11 g, 68%): MS (ESI+) for C₂₈H₄₈N₄O₅ m/z 521.30(M+H)⁺.; HPLC purity 95% (ret. time, 1.07 min); ¹H-NMR (500 MHz,Chloroform-d) δ 7.41 (s, 1H), 5.28 (dt, J=9.3, 4.2 Hz, 1H), 4.05 (d,J=10.1 Hz, 1H), 3.71-3.64 (m, 1H), 3.64 (s, 3H), 3.31 (d, J=31.3 Hz,4H), 2.83 (s, 3H), 2.66 (ddt, J=12.5, 7.4, 3.7 Hz, 1H), 2.46 (s, 2H),2.35-2.28 (m, 1H), 2.20 (s, 3H), 2.13-1.95 (m, 4H), 1.92-1.76 (m, 3H),1.75-1.49 (m, 5H), 1.44 (s, 9H), 1.03 (d, J=7.0 Hz, 6H).

Step 10: tert-butylN-{2-[({3-[(3S)-4,4-dimethyl-3-[(3-methylbutyl)carbamoyl]cyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate

To a solution of 2 M Me₃Al in toluene (230 μl, 0.46 mmol) was added asolution of 3-methylbutan-1-amine (53.5 μl, 0.46 mmol) in toluene (1 ml)in a sealed tube. After 5 min, a solution of methyl(1S)-5-(4-{[(2-{[(tert-butoxy)carbonyl](methyl)amino}ethyl)(methyl)amino]methyl}-1-(oxan-2-yl)-1H-pyrazol-3-yl)-2,2-dimethylcyclohexane-1-carboxylate(200 mg, 0.0.38 mmol) in toluene (3 ml) was added. The reaction wassealed and heated to 110° C. for 18 h. The reaction was allowed to coolto RT and MeOH (25 ml) was added. The mixture was stirred at RT withCelite (˜5 g) filtered and the pad washed with MeOH (10 ml). Thefiltrates were concentrated. The crude product was absorbed onto silicagel (2 ml). Purification by silica gel column chromatography, on aBiotage Isolera system, using a 10 g HP-Sil SNAP cartridge, eluting withTHF:heptanes (1:9-1), gave the desired product as a colourless glass(160 mg, 72%): MS (ESI+) for C₃₂H₅₇N₅O₄ m/z 576.3 (M+H)⁺.; HPLC purity100% (ret. time, 1.14 min); ¹H-NMR (500 MHz, Chloroform-d) δ 7.57-7.34(m, 1H), 5.84-5.39 (m, 1H), 5.28 (s, 1H), 4.05 (d, J=11.9 Hz, 1H),3.72-3.62 (m, 1H), 3.41-3.14 (m, 4H), 2.82 (s, 2H), 2.77-2.36 (m, 2H),2.18 (s, 2H), 2.03 (d, J=17.2 Hz, 5H), 1.92-1.73 (m, 2H), 1.66 (s, 3H),1.55 (s, 9H), 1.52-1.42 (m, 9H), 1.41-1.30 (m, 3H), 1.13-1.06 (m, 2H),1.03 (s, 3H), 0.93-0.86 (m, 6H).

Step 11:(1S)-2,2-dimethyl-5-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]-N-(3-methylbutyl)cyclohexane-1-carboxamide(Compound 263)

6 N HCl (2 ml) was added to a solution of tert-butylN-{2-[({3-[(3R)-4,4-dimethyl-3-[(3-methylbutyl)carbamoyl]cyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate(160 mg, 0.28 mmol) in 1,4-dioxane (1 ml) at 0° C. and stirred for 5min. This was then allowed to continue at RT for 18 hr beforeevaporating in vacuo. MeOH (10 ml) was added to the residue andevaporated to dryness again. MeOH (5 ml) was added to the residue andthis solution was passed through an Isolute SCX 2 cartridge (2 g)followed by MeOH (2×5 ml). The product was eluted with 7 N NH₃ in MeOH(15 ml). This was then evaporated to dryness to give 90 mg (83%, 8:1cis:trans mixture) the desired product as a colourless glass: MS (ESI+)for C₂₂H₄₁N₅O m/z 392.2 (M+H)⁺.; HPLC purity 100% (ret. time, 2.47 min);¹H-NMR (500 MHz, Methanol-d4) δ 7.43 (s, 1H), 3.42 (d, J=4.5 Hz, 2H),3.27-3.18 (m, 1H), 3.18-3.08 (m, 1H), 2.85-2.75 (m, 1H), 2.71 (t, J=6.5Hz, 2H), 2.52 (t, J=6.5 Hz, 2H), 2.44-2.35 (m, 3H), 2.24-2.13 (m, 4H),2.07 (q, J=12.8 Hz, 1H), 1.90-1.78 (m, 1H), 1.75-1.58 (m, 3H), 1.58-1.51(m, 1H), 1.49-1.31 (m, 3H), 1.11 (d, J=6.8 Hz, 3H), 1.00 (d, J=18.6 Hz,3H), 0.93 (s, 3H), 0.92 (s, 3H).

Compound 271(1S,5R)—N-(3-methoxypropyl)-2,2-dimethyl-5-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]cyclohexane-1-carboxamide

Step 1: tert-butylN-{2-[({3-[(3S)-3-[(3-methoxypropyl)carbamoyl]-4,4-dimethylcyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate

To a solution of 2 M Me₃Al in toluene (230 μl, 0.46 mmol) was added asolution of 3-methoxypropylamine (47.0 μl, 0.46 mmol) in toluene (1 ml)in a sealed tube. After 5 min, a solution of methyl(1S)-5-(4-{[(2-{[(tert-butoxy)carbonyl](methyl)amino}ethyl)(methyl)amino]methyl}-1-(oxan-2-yl)-1H-pyrazol-3-yl)-2,2-dimethylcyclohexane-1-carboxylate(200 mg, 0.38 mmol) in toluene (3 ml) was added. The reaction was sealedand heated to 110° C. for 18 h. The reaction was allowed to cool to RTand MeOH (25 ml) was added. The mixture was stirred at RT with Celite(˜5 g) and filtered and the pad washed with MeOH (10 ml). The filtrateswere concentrated. The crude product was absorbed onto silica gel (2ml). Purification by silica gel column chromatography, on a BiotageIsolera system, using a 25 g KP-Sil SNAP cartridge, eluting withTHF:heptanes (1:9-1), gave the desired product as a colourless glass(130 mg, 58%): MS (ESI+) for C₃₁H₅₅N₅O₅ m/z 578.35 (M+H)⁺.; HPLC purity100% (ret. time, 1.11 min); ¹H-NMR (500 MHz, Chloroform-d) δ 7.67-7.30(m, 1H), 5.37-5.18 (m, 1H), 4.04 (d, J=11.0 Hz, 1H), 3.67 (t, J=11.4 Hz,1H), 3.45 (t, J=5.5 Hz, 2H), 3.39-3.20 (m, 8H), 3.02-2.77 (m, 3H), 2.70(s, 1H), 2.46 (s, 2H), 2.19 (s, 2H), 2.10-1.96 (m, 5H), 1.85 (s, 1H),1.80-1.73 (m, 3H), 1.72-1.62 (m, 3H), 1.57 (s, 6H), 1.45 (s, 9H), 1.09(s, 3H), 1.06-1.01 (m, 3H).

Step 2:(1S,5R)—N-(3-methoxypropyl)-2,2-dimethyl-5-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]cyclohexane-1-carboxamide(Compound 271)

6 N HCl (2 ml) was added to a solution of tert-butylN-{2-[({3-[(3S)-3-[(3-methoxypropyl)carbamoyl]-4,4-dimethylcyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate(130 mg, 0.23 mmol) in 1,4-dioxane (1 ml) at 0° C. and stirred for 5min. This was then allowed to continue at RT for 18 hr beforeevaporating in vacuo. MeOH (10 ml) was added to the residue andevaporated to dryness again. MeOH (5 ml) was added to the residue andthis solution was passed through an Isolute SCX 2 cartridge (2 g)followed by MeOH (2×5 ml). The product was eluted with 7 N NH₃ in MeOH(15 ml). This was then evaporated to dryness to give 69 mg (78%) of thedesired product as a colourless glass. The diastereoisomers wereseparated by prep HPLC under high pH conditions to give 3 mg of thedesired product: MS (ESI+) for C₂₁H₃₉N₅O₂ m/z 394.5 (M+H)⁺.; HPLC purity91% (ret. time, 1.00 min); ¹H-NMR (500 MHz, Methanol-d4) δ 7.43 (s, 1H),3.69 (s, 1H), 3.51-3.39 (m, 4H), 3.32 (d, J=1.0 Hz, 3H), 3.31-3.24 (m,1H), 3.22-3.12 (m, 1H), 2.92 (t, J=5.8 Hz, 2H), 2.63-2.50 (m, 5H), 2.20(d, J=10.8 Hz, 4H), 2.15-1.99 (m, 2H), 1.87-1.65 (m, 5H), 1.38-1.27 (m,1H), 1.13 (s, 3H), 0.99 (s, 3H) (plus 51 mg of(1S,55S)—N-(3-methoxypropyl)-2,2-dimethyl-5-[4-({methyl[2-(methylamino)ethyl]amino}methyl)-1H-pyrazol-3-yl]cyclohexane-1-carboxamide).

Compound 273 ({3-[(3R)-4,4-dimethyl-3-(oxan-4-ylmethoxy)cyclohexyl]-1H-pyrazol-4-yl}methyl)(methyl)[2-(methylamino)ethyl]amine

Step 1: (1S,6S)-5,5-dimethyl-7-oxabicyclo[4.1.0]heptan-2-one

(1S,2S)-1,2-diphenylethane-1,2-diamine (3.42 g, 16.11 mmol) andtrifluoroacetic acid (1.2 ml, 16.11 mmol) were dissolved in 1,4-dioxane(150 ml). The solution was stirred for 30 min before adding4,4-dimethylcyclohex-2-en-1-one (10 g, 80.53 mmol) and hydrogen peroxide(10.58 ml, 120.79 mmol 35% in water). The reaction was stirred andheated to 50° C. for 72 h after which time the reaction was quenchedwith NH₄Cl (saturated, 100 ml). The solution was extracted with DCM(4×100 ml). The combined organic extracts were dried over Na₂SO₄ andevaporated to dryness to afford 12.5 g of desired material (containing−10% 1,4-dioxane w/w). ¹H-NMR (250 MHz, Chloroform-d) δ 3.23 (d, J=4.0Hz, 1H), 3.17 (dd, J=4.0, 1.2 Hz, 1H), 2.41 (ddd, J=18.8, 6.5, 3.2 Hz,1H), 2.19 (ddd, J=18.7, 11.5, 6.9 Hz, 1H), 1.90 (td, J=12.5, 11.5, 6.5Hz, 1H), 1.35 (dtd, J=9.9, 3.1, 1.2 Hz, 1H), 1.22 (s, 3H), 1.06 (s, 3H).Rf=0.30 (3% 7 N NH₃ in MeOH in DCM).

Step 2: (3R)-3-hydroxy-4,4-dimethylcyclohexan-1-one

At RT under nitrogen, lithium (1.63 g, 235 mmol) was added to a solutionof naphthalene (40 g, 314 mmol) in dry THF (600 ml). The solutionquickly turned dark green and the reaction was stirred at RT until fulldissolution of the lithium (˜5 h). The solution was cooled to −78° C.and a solution of (1S,6S)-5,5-dimethyl-7-oxabicyclo[4.1.0]heptan-2-one(11 g, 78.47 mmol) in dry THF (300 ml) was added. The reaction wasstirred for 1 h then quenched with water (30 ml) and allowed to warm toRT. A further 300 ml of water was added and the solution was extractedwith Et₂O (2×500 ml). The combined organic extracts were dried overNa₂SO₄ and evaporated to dryness. The residue was purified by Biotage(SNAP 340 g, eluent heptane/EtOAc/NEt₃ 90/10/1 to 10/90/1) to afford5.81 g of title compound (52%) as an orange oil. ¹H-NMR (500 MHz,Chloroform-d) δ 3.77-3.62 (m, 1H), 2.64 (ddd, J=14.9, 4.3, 1.0 Hz, 1H),2.46-2.36 (m, 1H), 2.36-2.25 (m, 2H), 1.94-1.82 (m, 1H), 1.83-1.76 (m,1H), 1.54-1.44 (m, 1H), 1.13 (s, 3H), 1.07 (s, 3H). Rf=0.30(EtOAc/heptane/NEt₃ (6/4/0.1).

Step 3:(3R)-3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohexan-1-one

(3R)-3-hydroxy-4,4-dimethylcyclohexan-1-one (5.81 g, 40.86 mmol),tert-butyl(chloro)dimethylsilane (9.24 g, 61.29 mmol) and 1H-imidazole(6.95 g, 102.15 mmol) were dissolved in DMF (50 ml). The reaction wasstirred at RT overnight; no starting material was detected by TLC. Thereaction was quenched with saturated aqueous ammonium chloride solution(30 ml) and was extracted with EtOAc (3×30 ml); the combined organiclayers were washed with water (30 ml) and dried over Na₂SO₄, evaporatedand co-evaporated with toluene (4×50 ml) to dryness affording 8.4 g ofthe title compound isolated as a yellow oil (80%). ¹H-NMR (500 MHz,Chloroform-d) δ 3.64 (dd, J=7.4, 4.1 Hz, 1H), 2.63-2.49 (m, 1H),2.39-2.25 (m, 3H), 1.95-1.78 (m, 1H), 1.43 (dt, J=13.8, 7.1 Hz, 1H),1.07 (s, 3H), 1.01 (s, 3H), 0.88 (s, 9H), 0.04 (d, J=6.0 Hz, 6H).Rf=0.53 (heptane/EtOAc 85/15).

Step 4:(3R)-3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohex-1-en-1-yltrifluoromethanesulfonate

(3R)-3-[(tert-Butyldimethylsilyl)oxy]-4,4-dimethylcyclohexan-1-one (3 g,11.7 mmol) was dissolved in dry THF (250 ml). The solution was cooled to−78° C. and 1 M lithium 1,1,1,3,3,3-hexamethyldisilazan-2-ide (23.4 ml)was slowly added. The reaction was stirred for 45 min and a solution ofN-(5-chloropyridin-2-yl)-1,1,1-trifluoro-N-[(trifluoromethyl)sulfonyl]methanesulfonamide(8.59 g, 21.88 mmol) in dry THF (60 ml) was slowly added. The reactionwas allowed to warm to RT and stirred for 3 h. The reaction was quenchedwith NH₄Cl (saturated, 100 ml) and extracted with EtOAc (3×100 ml). Thecombined organic extracts were dried over Na₂SO₄ and evaporated todryness and the residue purified by Biotage (SNAP HP 100 g, eluentheptane/EtOAc 100/0 to 90/10) to afford 3.1 g of title compound as a 1:1mix of isomers (61%). ¹H-NMR (500 MHz, Chloroform-d) δ 5.74-5.52 (m,1H), 3.57 (t, J=5.3 Hz, 1H), 2.53 (dd, J=17.3, 2.2 Hz, 1H), 2.32-2.21(m, 1H), 2.13 (ddt, J=17.6, 4.4, 2.5 Hz, 1H), 1.88 (ddt, J=17.5, 4.4,2.4 Hz, 1H), 0.96-0.84 (m, 15H), 0.06 (d, J=7.4 Hz, 6H).

Step 5:tert-butyl({[(1R)-6,6-dimethyl-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-en-1-yl]oxy})dimethylsilane

A suspension of(3R)-3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohex-1-en-1-yltrifluoromethanesulfonate (90%, 3.11 g, 7.2 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi-1,3,2-dioxaborolane (2.74 g,10.81 mmol) and potassium acetate (3.15 ml, 50.43 mmol) in 1,4-dioxane(100 ml) was degassed with a N₂ sparge for 10 min whilst stirring at RT.Bis[3-(diphenylphosphanyl)cyclopenta-2,4-dien-1-yl]iron;dichloromethane; dichloropalladium (0.59 g, 0.72 mmol) was added to thissuspension and stirred at 90° C. for 3.5 h before allowing to cool to RTovernight. The reaction mixture was diluted with EtOAc (50 ml) and water(50 ml). The organic layer was separated and the aqueous was extractedwith EtOAc (3×50 ml). The combined organic layers were dried overNa₂SO₄, filtered and evaporated in vacuo. Purification by silica gelcolumn chromatography, on a Biotage Isolera system, using a 100 g HP-SilSNAP cartridge, eluting with EtOAc:heptanes (0-5:95), gave the desiredproduct as an orange oil (2.02 g, 76%). ¹H-NMR (500 MHz, Chloroform-d) δ6.50-6.12 (m, 1H), 3.90-3.41 (m, 1H), 2.34-2.18 (m, 1H), 2.18-1.96 (m,2H), 1.96-1.82 (m, 1H), 1.25 (d, J=3.2 Hz, 12H), 0.95-0.80 (m, 15H),0.11-−0.01 (m, 6H).

Step 6: tert-butylN-{2-[({3-[(3R)-3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohex-1-en-1-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate

tert-Butyl({[(1R)-6,6-dimethyl-3-(tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-2-en-1-yl]oxy})dimethylsilane (2 g, 5.46 mmol), tert-butylN-[2-({[3-iodo-1-(oxan-2-yl)-1H-pyrazol-4-yl]methyl}(methyl)amino)ethyl]-N-methylcarbamate(2.61 g, 5.46 mmol) and potassium carbonate (2.26 g, 16.37 mmol) weresuspended in 1,4-dioxane/water (240 ml, 7/1). The solution was degassedwith nitrogen for 10 min and Pd(dppf)Cl₂ (0.45 g, 0.55 mmol) was added.The reaction was heated to 100° C. After overnight, the solvents wereevaporated. The residue was purified by Biotage (SNAP HP 100 g, eluentheptane/EtOAc (+1% NEt₃) 95/5 to 60/40) to afford 2.5 g of the titlecompound as a yellow oil (62%; at 80% purity). ¹H-NMR (500 MHz,Chloroform-d) δ 7.61-7.36 (m, 1H), 6.14-5.72 (m, 1H), 5.43-5.23 (m, 1H),4.08-3.92 (m, 1H), 3.76-3.57 (m, 1H), 3.51-3.19 (m, 5H), 2.99-2.62 (m,6H), 2.60-2.30 (m, 3H), 2.28-2.14 (m, 3H), 2.13-1.94 (m, 4H), 1.76-1.51(m, 6H), 1.51-1.37 (m, 9H), 1.00-0.79 (m, 18H), 0.13-−0.03 (m, 6H).Rf=0.29 (EtOAc/heptane 7/3+1% NEt₃).

Step 7: tert-butylN-{2-[({3-[(3R)-3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate

A solution of tert-butylN-{2-[({3-[(3R)-3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohex-1-en-1-yl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate(80%, 1 g, 1.35 mmol) in EtOH (10 ml) was cautiously added onto a purged[nitrogen] suspension of Raney-Nickel catalyst (2.5 ml) in EtOH (20 ml).The resulting solution was purged with nitrogen (3×), hydrogen (2×) andleft under an atmosphere of hydrogen at RT. After overnight, an aliquotwas analysed showing only starting material. Additional 7.5 ml ofcatalyst was added and the reaction was left stirring under hydrogenatmosphere for 6 h, after which time LCMS showed complete conversion tothe desired product. The solution was filtered through Celite and thepad was washed with EtOAc (150 ml). The filtrate was evaporated underreduced pressure and co-evaporated with toluene to afford 870 mg of thetitle compound as light yellow oil (92%). ¹H-NMR (250 MHz, Chloroform-d)δ 7.45 (d, J=11.3 Hz, 1H), 5.41-5.17 (m, 1H), 4.06 (d, J=8.3 Hz, 1H),3.67 (t, J=11.2 Hz, 1H), 3.34 (d, J=8.8 Hz, 6H), 2.85 (d, J=12.0 Hz,5H), 2.68 (s, 1H), 2.47 (s, 2H), 2.20 (d, J=9.2 Hz, 3H), 2.02 (s, 3H),1.93-1.36 (m, 13H), 1.25 (d, J=6.2 Hz, 2H), 1.06-0.79 (m, 15H), 0.02 (d,J=7.0 Hz, 6H).

Step 8: tert-butylN-{2-[({3-[(3R)-3-hydroxy-4,4-dimethylcyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate

tert-ButylN-{2-[({3-[(3R)-3-[(tert-butyldimethylsilyl)oxy]-4,4-dimethylcyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate(85%, 870 mg, 1.25 mmol) was dissolved in 1 M TBAF in THF (12 ml). Thereaction was heated to 60° C. and stirred overnight. The reaction wasquenched with water (10 ml) and was extracted with EtOAc (3×20 ml). Thecombined organic extracts were dried over Na₂SO₄ and evaporated and todryness. The residue was purified by Biotage (SNAP 50 g, eluent DCM/MeOH100/0 to 90/10) to afford 450 mg of title compound as a light yellow oil(60%). ¹H-NMR (250 MHz, Chloroform-d) δ 7.53-7.31 (m, 1H), 5.30-5.13 (m,1H), 3.98 (d, J=10.3 Hz, 1H), 3.60 (td, J=11.1, 2.8 Hz, 1H), 3.31 (dd,J=12.4, 6.8 Hz, 9H), 2.89 (dd, J=10.3, 6.5 Hz, 1H), 2.78 (d, J=13.1 Hz,9H), 2.50-2.29 (m, 2H), 2.12 (s, 3H), 1.91 (d, J=20.2 Hz, 4H), 1.77-1.47(m, 8H), 1.37 (d, J=7.5 Hz, 26H), 1.24-1.12 (m, 2H), 1.04-0.83 (m, 8H).Rf=0.14 (DCM/MeOH 95/5).

Step 9: tert-butylN-{2-[({3-[(3R)-4,4-dimethyl-3-(oxan-4-ylmethoxy)cyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate

Potassium hexamethyldisilazide (3.44 ml, 0.91 M in THF) and 18-crown-6(17 mg, 0.06 mmol) were added to a solution of tert-butylN-{2-[({3-[(3R)-3-hydroxy-4,4-dimethylcyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate(300 mg, 0.63 mmol) in dry toluene (10 ml). The reaction was stirred atRT for 1 h, then 4-(bromomethyl)tetrahydro-2H-pyran (250 μl, 1.88 mmol)was added and the solution was heated to 70° C. while monitoring byLCMS. Further aliquots of potassium hexamethyldisilazide (1.5 ml, 0.91 Min THF) and 4-(bromomethyl)tetrahydro-2H-pyran (100 μl, 0.75 mmol) wereadded after 16 h, 24 h, 48 h and 72 h. The reaction was stopped after 1week. The solution was washed with water (25 ml) and extracted withEtOAc (3×30 ml). The combined organic extracts were dried over Na₂SO₄and evaporated to dryness. The crude residue was purified by low pH prepHPLC in three injections; the product rich fractions were combined(co-evaporated with toluene) to afford 7 mg of desired alkylated (2%).60 mg starting material were also recovered. ¹H-NMR (500 MHz,Chloroform-d) δ 7.89-7.43 (m, 1H), 5.40-5.20 (m, 1H), 4.15-3.86 (m, 3H),3.81-3.62 (m, 2H), 3.61-3.31 (m, 7H), 3.16-3.05 (m, 1H), 2.97-2.71 (m,5H), 2.68-2.50 (m, 2H), 2.48-2.22 (m, 3H), 2.13-1.92 (m, 4H), 1.89-1.18(m, 21H), 1.02 (s, 3H), 0.95 (s, 3H). LC-MS: 1.24 min (2.5 minute LC-MSmethod), m/z=577.35.

Step 10: ({3-[(3R)-4,4-dimethyl-3-(oxan-4-ylmethoxy)cyclohexyl]-1H-pyrazol-4-yl}methyl)(methyl)[2-(methylamino)ethyl]amine(Compound 273)

tert-ButylN-{2-[({3-[(3R)-4,4-dimethyl-3-(oxan-4-ylmethoxy)cyclohexyl]-1-(oxan-2-yl)-1H-pyrazol-4-yl}methyl)(methyl)amino]ethyl}-N-methylcarbamate(7 mg, 0.01 mmol) was dissolved in 1,4-dioxane (2 ml) and HCl (6 N, 1ml) was added. After 2 h stirring at RT, the solvent was removed underreduced pressure to afford 4 mg the title compound (84%). ¹H-NMR (500MHz, Methanol-d4) δ 8.44 (s, 1H), 4.68-4.43 (m, 2H), 3.92 (d, J=11.5 Hz,2H), 3.84-3.57 (m, 4H), 3.51 (dd, J=8.8, 6.2 Hz, 1H), 3.46-3.37 (m, 2H),3.28-3.16 (m, 2H), 2.93 (d, J=4.3 Hz, 3H), 2.80 (s, 3H), 2.24-2.13 (m,1H), 1.87-1.48 (m, 8H), 1.41-1.25 (m, 2H), 1.06 (d, J=2.6 Hz, 3H), 1.00(s, 3H). LC-MS: 2.47 min (7 min method), m/z=393.2.

Compound 274N¹-((4-(4-fluorophenyl)isoxazol-5-yl)methyl)-N¹-methylethane-1,2-diaminetrifluoroacetate

Step 1:3,3-dimethyl-1-oxaspiro[4.5]decan-8-ylidene](tert-butoxy)carbohydrazide

Into a 50-mL round-bottom flask, was placed hexane (10 mL),(tert-butoxy)carbohydrazide (2.64 g, 19.98 mmol, 1.00 equiv), and3,3-dimethyl-1-oxaspiro[4.5]decan-8-one (3.65 g, 20.03 mmol, 1.00equiv). The resulting solution was stirred for 15 h at 75° C. and thenallowed to cool to room temperature. The solids were collected byfiltration to give 4 g (67%) of3,3-dimethyl-1-oxaspiro[4.5]decan-8-ylidene](tert-butoxy)carbohydrazideas a white solid. ¹H-NMR (300 MHz, DMSO-d6): δ 9.52 (s, 1H), 3.43 (s,2H), 2.48-2.09 (m, 4H), 1.86-1.46 (m, 6H), 1.42 (s, 9H), 1.06 (s, 6H)ppm.

Step 2:3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl](tert-butoxy)carbohydrazide

Into a 250-mL round-bottom flask, was placed3,3-dimethyl-1-oxaspiro[4.5]decan-8-ylidene](tert-butoxy)carbohydrazide(3.5 g, 11.81 mmol, 1.00 equiv), ethanol (60 mL), and 10%palladium/carbon (0.35 g). Hydrogen (3 atm) was then applied to thereaction mixture. The reaction mixture was stirred for 48 h at roomtemperature. The solids were filtered and the solution was concentratedunder vacuum. This resulted in 3.5 g (99%) of3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl](tert-butoxy)carbohydrazide as awhite solid. ¹H-NMR (300 MHz, DMSO-d6): δ 8.15 (s, 1H), 4.12 (s, 1H),3.36 (s, 2H), 2.79-2.56 (m, 1H), 1.80-1.60 (m, 2H), 1.46-1.34 (m, 3H),1.34-1.20 (m, 14H), 1.02 (s, 6H) ppm.

Step 3: [3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]hydrazine hydrochloridesalts

Into a 100-mL round-bottom flask, was placed3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl](tert-butoxy)carbohydrazide (3.5g, 11.73 mmol, 1.00 equiv) and a solution of saturated hydrogen chloridegas in methanol (35 mL). The resulting solution was stirred for 15 h atroom temperature and then concentrated under vacuum. This resulted in2.7 g (98%) of [3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]hydrazinehydrochloride salts as a white solid. ¹H-NMR (300 MHz, D₂O): δ 3.43 (s,2H), 3.18-3.00 (m, 1H), 2.10-1.75 (m, 4H), 1.75-1.25 (m, 6H), 1.00 (s,6H) ppm.

Step 4:1-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazole-5-carbaldehyde

Into a 100-mL round-bottom flask, was placed[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]hydrazine hydrochloride (1.0 g,4.26 mmol, 1.00 equiv),[(1E)-4,4-dimethoxy-3-oxobut-1-en-1-yl]dimethylamine (740 mg, 4.27 mmol,1.00 equiv), and methanol (25 mL). The resulting solution was stirredfor 15 h at 70° C. and then concentrated under vacuum. The residue wasdiluted with THF (10 mL) and hydrochloric acid (1N, 15 mL) and thenstirred at room temperature for 2 h. The THF was removed under vacuumand the residue was extracted with 3×30 mL of ethyl acetate and theorganic layers combined and dried over anhydrous sodium sulfate andconcentrated under vacuum. The residue was purified by silica gel columnwith ethyl acetate/petroleum ether (2:3). This resulted in 250 mg (22%)of 1-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazole-5-carbaldehydeas a white solid. ¹H-NMR (300 MHz, CDCl₃): δ 9.85 (s, 1H), 7.54 (d,J=2.1 Hz, 1H), 6.87 (d, J=2.1 Hz, 1H), 5.05-4.90 (m, 1H), 3.51 (s, 2H),2.45-2.25 (m, 2H), 2.05-1.90 (m, 2H), 1.90-1.75 (m, 2H), 1.65-1.50 (m,4H), 1.10 (s, 6H) ppm.

Step 5: tert-butylN-(2-[[(1-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate

Into a 250-mL round-bottom flask, was placed1-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazole-5-carbaldehyde(524 mg, 2.00 mmol, 1.00 equiv), tert-butylN-methyl-N-[2-(methylamino)ethyl]carbamate (590 mg, 3.13 mmol, 1.57equiv), ClCH₂CH₂Cl (50 mL). Then NaBH(OAc)₃ (3.39 g, 16.00 mmol, 8.01equiv) was added by batchwise at 0° C. The resulting solution wasstirred for 3 h at 0° C. The reaction was then quenched by the additionof 50 mL of Na₂CO₃ (sat. aq.). The resulting solution was extracted with3×50 mL of ethyl acetate and the organic layers combined and dried overanhydrous sodium sulfate and concentrated under vacuum. The residue wasapplied onto a C18 gel column with CH₃CN/H₂O (4:1). This resulted in 650mg (75%) of tert-butylN-(2-[[(1-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl](methyl)amino]ethyl)-N-methylcarbamateas light yellow oil.

Step 6:N¹-((4-(4-fluorophenyl)isoxazol-5-yl)methyl)-N¹-methylethane-1,2-diaminetrifluoroacetic acid (Compound 274)

Into a 50-mL round-bottom flask, was placed tert-butylN-(2-[[(1-[3,3-dimethyl-1-oxaspiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl](methyl)amino]ethyl)-N-methylcarbamate(600 mg, 1.38 mmol, 1.00 equiv) and ta solution of saturated hydrogenchloride gas in methanol (6 mL). The resulting solution was stirred for3 h at room temperature and then concentrated under vacuum. The crudeproduct was purified by Prep-HPLC with the following conditions(Prep-HPLC-025): Column, XBridge Shield RP 18 OBD Column, 5 μm, 19*150mm, mobile phase, Water with 10 mmol TFA and MeCN (5.0% MeCN up to 21.0%in 6 min, 21.0% 7 min); Detector, UV 254/220 nm. This resulted in 550 mg(71%) ofN¹-((4-(4-fluorophenyl)isoxazol-5-yl)methyl)-N¹-methylethane-1,2-diaminetrifluoroacetic acid salt as a colorless oil. ¹H-NMR (300 MHz, CDCl₃):δ7.60 (d, J=1.8 Hz, 1H), 6.53 (d, J=1.8 Hz, 1H), 4.57 (s, 2H), 4.32-4.15(m, 1H), 3.61-3.41 (m, 2H), 2.81 (s, 3H), 2.70 (s, 3H), 2.15-1.85 (m,4H), 1.77-1.45 (m, 6H), 0.98 (s, 6H) ppm. LCMS (method A, ESI): RT=4.73min, m/z=335 [M+H]⁺.

Compound 275 Methyl[2-(methylamino)ethyl][(1-[spiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl]aminetrifluoroacetate

Step 1: spiro[4.5]decan-8-ylidene (tert-butoxy)carbohydrazide

Into a 100-mL round-bottom flask, was placed spiro[4.5]decan-8-one (1.52g, 9.98 mmol, 1.00 equiv), (tert-butoxy)carbohydrazide (1.32 g, 9.99mmol, 1.00 equiv), hexane (20 mL). The resulting solution was stirredfor 12 h at 75° C. then cooled to room temperature and concentratedunder vacuum. The residue was triturated with 1×5 mL of hexane and thesolids were collected by filtration to afford 2.13 g (80%) ofspiro[4.5]decan-8-ylidene (tert-butoxy)carbohydrazide as a white solid.¹H-NMR (300 MHz, DMSO-d6): δ 9.49 (s, 1H), 2.29 (d, J=6.3 Hz, 2H), 2.18(d, J=6.3 Hz, 2H), 1.67-1.32 (m, 21H) ppm.

Step 2: spiro[4.5]decan-8-ylhydrazine hydrochloride

To a solution of spiro[4.5]decan-8-ylidene (tert-butoxy)carbohydrazide(2 g, 7.51 mmol, 1.00 equiv) in tetrahydrofuran (10 mL) at −40° C. underdry nitrogen was added a solution of borane (1 M in THF; 8.3 mL, 1.10equiv) dropwise over approximately 20 min. The resulting solution wasstirred for 1 h at room temperature then treated dropwise withhydrochloric acid (6 N, 5 mL) with stirring. The resulting solution wasstirred for 12 h at room temperature and then concentrated under vacuum.The residue was triturated with 1×50 mL of ether and the solids werecollected by filtration to afford 2.5 g (crude) ofspiro[4.5]decan-8-ylhydrazine hydrochloride as a white solid. ¹H-NMR(300 MHz, DMSO-d6): δ2.94-2.80 (m, 1H), 1.95-1.80 (m, 2H), 1.45-1.45 (m,6H), 1.45-1.29 (m, 6H), 1.29-1.15 (m, 2H) ppm.

Step 3: 1-[spiro[4.5]decan-8-yl]-1H-pyrazole-5-carbaldehyde

Into a 100-mL round-bottom flask, was placedspiro[4.5]decan-8-ylhydrazine hydrochloride (1.64 g, 8.01 mmol, 1.00equiv), [(1E)-4,4-dimethoxy-3-oxobut-1-en-1-yl]dimethylamine (2.08 g,12.01 mmol, 1.50 equiv), methanol (40 mL). The resulting solution wasstirred for 12 h at 75° C. then cooled to room temperature andconcentrated under vacuum. The residue was diluted with 10 mL ofhydrochloric acid (6 N) and 30 mL THF and the resulting solution stirredfor 2 h at room temperature. The resulting mixture was extracted with3×50 mL of ethyl acetate and the combined organic layers dried overanhydrous sodium sulfate and concentrated under vacuum. The residue waspurified by flash chromatography on silica gel column using ethylacetate/petroleum ether (1:10) as eluent to afford 160 mg (9%) of1-[spiro[4.5]decan-8-yl]-1H-pyrazole-5-carbaldehyde as a light yellowoil. ¹H-NMR (300 MHz, CDCl₃): δ 9.86 (s, 1H), 7.56 (d, J=1.8 Hz, 1H),6.89 (d, J=1.8 Hz, 1H), 5.05-4.90 (m, 1H), 2.16-1.95 (m, 2H), 1.95-1.80(m, 2H), 1.75-1.35 (m, 12H) ppm.

Step 4: tert-butylN-methyl-N-(2-[methyl[(1-[spiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl]amino]ethyl)carbamate

To a solution of 1-[spiro[4.5]decan-8-yl]-1H-pyrazole-5-carbaldehyde(210 mg, 0.90 mmol, 1.00 equiv) and tert-butylN-methyl-N-[2-(methylamino)ethyl]carbamate (340 mg, 1.81 mmol, 2.00equiv) in DCE (20 mL) was added NaBH(OAc)₃ (1.54 g, 7.26 mmol, 8.04equiv) portionwise. The resulting solution was stirred for 12 h at roomtemperature then quenched with 20 mL of sodium carbonate (sat. aq.). Theorganic layer was collected and the aqueous layer was extracted with2×20 mL of ethyl acetate and the organic layers combined. The combinedorganics were dried over anhydrous sodium sulfate and concentrated undervacuum. The residue was purified by flash chromatography on silica gelcolumn using ethyl acetate/petroleum ether (1:1) as eluent to afford 240mg (66%) of tert-butylN-methyl-N-(2-[methyl[(1-[spiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl]amino]ethyl)carbamateas a light yellow oil. ¹H-NMR (300 MHz, CDCl₃): δ7.44 (s, 1H), 6.08 (s,1H), 4.28-4.11 (m, 1H), 3.50 (s, 2H), 3.42-3.20 (m, 2H), 2.83 (s, 3H),2.61-2.41 (m, 2H), 2.23 (s, 3H), 2.12-1.98 (m, 2H), 1.85-1.72 (m, 2H),1.70-1.51 (m, 6H), 1.51-1.32 (m, 15H) ppm.

Step 5:methyl[2-(methylamino)ethyl][(1-[spiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl]aminetrifluoroacetate (Compound 275)

Into a 50-mL round-bottom flask, was placed tert-butylN-methyl-N-(2-[methyl[(1-[spiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl]amino]ethyl)carbamate(210 mg, 0.52 mmol, 1.00 equiv) which was then dissolved in a solutionof saturated hydrogen chloride gas in 1,4-dioxane (10 mL). The reactionmixture was stirred for 12 h at room temperature and the resultantprecipitate was collected by filtration. The crude product was purifiedby Prep-HPLC with the following conditions (Prep-HPLC-025): Column,XBridge Shield RP18 OBD Column, 5 μm, 19×150 mm, mobile phase, Waterwith 10 mmol TFA and MeCN (5.0% MeCN up to 36.0% in 10 min); Detector,UV 254/220 nm. This resulted in 200 mg (72%) ofmethyl[2-(methylamino)ethyl][(1-[spiro[4.5]decan-8-yl]-1H-pyrazol-5-yl)methyl]aminetrifluoroacetate as a white semi-solid. ¹H-NMR (300 MHz, D₂O): δ 7.58(d, J=2.1 Hz, 1H), 6.50 (d, J=2.1 Hz, 1H), 4.54 (s, 2H), 4.22-4.16 (m,1H), 3.61-3.36 (m, 8H), 2.88 (s, 3H), 2.67 (s, 3H), 1.98-1.75 (m, 2H),1.74-1.57 (m, 2H), 1.57-1.20 (m, 12H). LCMS (method A, ESI): RT=1.09min, m/z=305.4 [M+1]⁺.

Biological Methods PRMT1 Biochemical Assay

General Materials.

S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), bicine,Tween20, dimethylsulfoxide (DMSO), bovine skin gelatin (BSG), andTris(2-carboxyethyl)phosphine hydrochloride solution (TCEP) werepurchased from Sigma-Aldrich at the highest level of purity possible.³H-SAM was purchase from American Radiolabeled Chemicals with a specificactivity of 80 Ci/mmol. 384-well streptavidin Flashplates were purchasedfrom PerkinElmer.

Substrates.

Peptide representative of human histone H4 residues 36-50 wassynthesized with an N-terminal linker-affinity tag motif and aC-terminal amide cap by 21^(st) Century Biochemicals. The peptide waspurified by high-performance liquid chromatography (HPLC) to greaterthan 95% purity and confirmed by liquid chromatography mass spectrometry(LC-MS). The sequence was Biot-Ahx-RLARRGGVKRISGLI-amide (SEQ ID NO.:1).

Molecular Biology:

Full-length human PRMT1 isoform 1 (NM_001536.5) transcript clone wasamplified from an HEK 293 cDNA library, incorporating flanking 5′sequence encoding a FLAG tag (DYKDDDDK) (SEQ ID NO.:2) fused directly toMet 1 of PRMT1. The amplified gene was subcloned into pFastBacI (LifeTechnologies) modified to encode an N-terminal GST tag and a TEVcleavage sequence(MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDENLYF QGGNS)(SEQ IDNO.:3) fused to Asp of the Flag tag of PRMT1.

Protein Expression.

Recombinant baculovirus were generated according to Bac-to-Bac kitinstructions (Life Technologies). Protein over-expression wasaccomplished by infecting exponentially growing High Five insect cellculture at 1.5×10⁶ cell/ml with 1:100 ratio of virus. Infections werecarried out at 27° C. for 48 hours, harvested by centrifugation, andstored at −80° C. for purification.

Protein Purification.

Expressed full-length human GST-tagged PRMT1 protein was purified fromcell paste by glutathione sepharose affinity chromatography afterequilibration of the resin with 50 mM phosphate buffer, 200 mM NaCl, 5%glycerol, 5 mM 3-mercaptoethanol, pH7.8 (Buffer A). GST-tagged PRMT1 waseluted with 50 mM Tris, 2 mM glutathione, pH 7.8, dialysed in buffer Aand concentrated to 1 mg/mL. The purity of recovered protein was 73%.Reference: Wasilko, D. J. and S. E. Lee: “TIPS: titerless infected-cellspreservation and scale-up” Bioprocess J., 5 (2006), pp. 29-32.

Predicted Translations:

GST-tagged PRMT1 (SEQ ID NO.: 4)MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDENLYFQGGNSDYKDDDDKMAAAEAANCIMENFVATLANGMSLQPPLEEVSCGQAESSEKPNAEDMTSKDYYFDSYAHFGIHEEMLKDEVRTLTYRNSMFHNRHLFKDKVVLDVGSGTGILCMFAAKAGARKVIGIECSSISDYAVKIVKANKLDHVVTIIKGKVEEVELPVEKVDIIISEWMGYCLFYESMLNTVLYARDKWLAPDGLIFPDRATLYVTAIEDRQYKDYKIHWWENVYGFDMSCIKDVAIKEPLVDVVDPKQLVTNACLIKEVDIYTVKVEDLTFTSPFCLQVKRNDYVHALVAYFNIEFTRCHKRTGFSTSPESPYTHWKQTVFYMEDYLTVKTGEEIFGTIGMRPNAKNNRDLDFTIDLDFKGQLCEL SCSTDYRMR

General Procedure for PRMT1 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween 20, prepared on theday of use. Compounds in 100% DMSO (1 ul) were spotted into apolypropylene 384-well V-bottom plates (Greiner) using a Platemate Plusoutfitted with a 384-channel head (Thermo Scientific). DMSO (1 ul) wasadded to Columns 11, 12, 23, 24, rows A-H for the maximum signal controland 1 ul of SAH, a known product and inhibitor of PRMT1, was added tocolumns 11, 12, 23, 24, rows I-P for the minimum signal control. Acocktail (40 ul) containing the PRMT1 enzyme was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate with PRMT1for 30 min at room temperature, then a cocktail (10 ul) containing SAMand peptide was added to initiate the reaction (final volume=51 ul). Thefinal concentrations of the components were as follows: PRMT1 was 0.5nM, ³H-SAM was 200 nM, non-radiolabeled SAM was 1.5 uM, peptide was 20nM, SAH in the minimum signal control wells was 1 mM, and the DMSOconcentration was 2%. The assays were stopped by the addition ofnon-radiolabeled SAM (10 ul) to a final concentration of 300 uM, whichdilutes the ³H-SAM to a level where its incorporation into the peptidesubstrate is no longer detectable. 50 ul of the reaction in the 384-wellpolypropylene plate was then transferred to a 384-well Flashplate andthe biotinylated peptides were allowed to bind to the streptavidinsurface for at least 1 hour before being washed once with 0.1% Tween20in a Biotek ELx405 plate washer. The plates were then read in aPerkinElmer TopCount plate reader to measure the quantity of ³H-labeledpeptide bound to the Flashplate surface, measured as disintegrations perminute (dpm) or alternatively, referred to as counts per minute (cpm).

% Inhibition Calculation

${\% \mspace{14mu} {inh}} = {100 - {( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} ) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{( {{Top} - {Bottom}} )}{( {1 + ( \frac{X}{{IC}_{50}} )^{{Hill}\mspace{14mu} {Coefficient}}} }}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

PRMT6 Biochemical Assay

General Materials.

S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), bicine,Tween20, dimethylsulfoxide (DMSO), bovine skin gelatin (BSG), sodiumbutyrate and Tris(2-carboxyethyl)phosphine hydrochloride solution (TCEP)were purchased from Sigma-Aldrich at the highest level of puritypossible. ³H-SAM was purchase from American Radiolabeled Chemicals witha specific activity of 80 Ci/mmol. 384-well streptavidin Flashplateswere purchased from PerkinElmer.

Substrates.

Peptide representative of human histone H4 residues 36-50 wassynthesized with an N-terminal linker-affinity tag motif and aC-terminal amide cap by 21st Century Biochemicals. The peptide waspurified by high-performance liquid chromatography (HPLC) to greaterthan 95% purity and confirmed by liquid chromatography mass spectrometry(LC-MS). The sequence was Biot-Ahx-RLARRGGVKRISGLI-amide and contained amonomethylated lysine at position 44 (SEQ ID NO.:5).

Molecular Biology:

Full-length human PRMT6 (NM_018137.2) transcript clone was amplifiedfrom an HEK 293 cDNA library, incorporating a flanking 5′ sequenceencoding a FLAG tag (MDYKDDDDK) (SEQ ID NO.:6) fused directly to Ser 2of PRMT6 and a 3′ sequence encoding a hexa His sequence (HHHHHH) (SEQ IDNO.:17) fused directly to Asp 375. The amplified gene was subcloned intopFastBacMam (Viva Biotech).

Protein Expression.

Recombinant baculovirus were generated according to Bac-to-Bac kitinstructions (Life Technologies). Protein over-expression wasaccomplished by infecting exponentially growing HEK 293F cell culture at1.3×10⁶ cell/ml with virus (MOI=10) in the presence of 8 mM sodiumbutyrate. Infections were carried out at 37° C. for 48 hours, harvestedby centrifugation, and stored at −80° C. for purification.

Protein Purification.

Expressed full-length human Flag- and His-tagged PRMT6 protein waspurified from cell paste by NiNTA agarose affinity chromatography afterequilibration of the resin with buffer containing 50 mM Tris, 300 mMNaCl, 10% glycerol, pH 7.8 (Buffer Ni-A). Column was washed with 20 mMimidazole in the same buffer and Flag-PRMT6-His was eluted with 150 mMimidazole. Pooled fractions were dialysed against buffer Ni-A andfurther purified by anti-flag M2 affinity chromatography. Flag-PRMT6-Hiswas eluted with 200 ug/ml FLAG peptide in the same buffer. Pooledfractions were dialysed in 20 mM Tris, 150 mM NaCl, 10% glycerol and 5mM j3-mercaptoethanol, pH 7.8. The purity of recovered protein was 95%.

Predicted Translations:

Flag-PRMT6-His (SEQ ID NO.: 7)MDYKDDDDKSQPKKRKLESGGGGEGGEGTEEEDGAEREAALERPRRTKRERDQLYYECYSDVSVHEEMIADRVRTDAYRLGILRNWAALRGKTVLDVGAGTGILSIFCAQAGARRVYAVEASAIWQQAREVVRFNGLEDRVHVLPGPVETVELPEQVDAIVSEWMGYGLLHESMLSSVLHARTKWLKEGGLLLPASAELFIAPISDQMLEWRLGFWSQVKQHYGVDMSCLEGFATRCLMGHSEIVVQGLSGEDVLARPQRFAQLELSRAGLEQELEAGVGGRFRCSCYGSAPMHGFAIWFQVTFPGGESEKPLVLSTSPFHPATHWKQALLYLNEPVQVEQDTDVSGEITLLPSRDNPRRLRVLLRYKVGDQEEKTKDFAMEDHHHHHH

General Procedure for PRMT6 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween 20, prepared on theday of use. Compounds in 100% DMSO (1 ul) were spotted into apolypropylene 384-well V-bottom plates (Greiner) using a Platemate Plusoutfitted with a 384-channel head (Thermo Scientific). DMSO (1 ul) wasadded to Columns 11, 12, 23, 24, rows A-H for the maximum signal controland 1 ul of SAH, a known product and inhibitor of PRMT6, was added tocolumns 11, 12, 23, 24, rows I-P for the minimum signal control. Acocktail (40 ul) containing the PRMT6 enzyme was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate with PRMT6for 30 min at room temperature, then a cocktail (10 ul) containing SAMand peptide was added to initiate the reaction (final volume=51 ul). Thefinal concentrations of the components were as follows: PRMT6 was 1 nM,³H-SAM was 200 nM, non-radiolabeled SAM was 250 nM, peptide was 75 nM,SAH in the minimum signal control wells was 1 mM, and the DMSOconcentration was 2%. The assays were stopped by the addition ofnon-radiolabeled SAM (10 ul) to a final concentration of 400 uM, whichdilutes the ³H-SAM to a level where its incorporation into the peptidesubstrate is no longer detectable. 50 ul of the reaction in the 384-wellpolypropylene plate was then transferred to a 384-well Flashplate andthe biotinylated peptides were allowed to bind to the streptavidinsurface for at least 1 hour before being washed once with 0.1% Tween20in a Biotek ELx405 plate washer. The plates were then read in aPerkinElmer TopCount plate reader to measure the quantity of ³H-labeledpeptide bound to the Flashplate surface, measured as disintegrations perminute (dpm) or alternatively, referred to as counts per minute (cpm).

% Inhibition Calculation

${\% \mspace{14mu} {inh}} = {100 - {( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} ) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{( {{Top} - {Bottom}} )}{( {1 + ( \frac{X}{{IC}_{50}} )^{{Hill}\mspace{14mu} {Coefficient}}} }}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

PRMT8 Biochemical Assay

General Materials.

S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), bicine,Tween20, dimethylsulfoxide (DMSO), bovine skin gelatin (BSG),isopropyl-β-D-thiogalactopyranoside (IPTG), andTris(2-carboxyethyl)phosphine hydrochloride solution (TCEP) werepurchased from Sigma-Aldrich at the highest level of purity possible.³H-SAM was purchase from American Radiolabeled Chemicals with a specificactivity of 80 Ci/mmol. 384-well streptavidin Flashplates were purchasedfrom PerkinElmer.

Substrates.

Peptide representative of human histone H4 residues 31-45 wassynthesized with an N-terminal linker-affinity tag motif and aC-terminal amide cap by 21^(st) Century Biochemicals. The peptide waspurified by high-performance liquid chromatography (HPLC) to greaterthan 95% purity and confirmed by liquid chromatography mass spectrometry(LC-MS). The sequence was Biot-Ahx-KPAIRRLARRGGVKR-amide (SEQ ID NO.:8).

Molecular Biology:

Full-length human PRMT8 (NM_019854.4) isoform 1 transcript clone wasamplified from an HEK 293 cDNA library and subcloned into pGEX-4T-1 (GELife Sciences). The resulting construct encodes an N-terminal GST tagand a thrombin cleavage sequence(MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGV SRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLVPRG SPEF) (SEQ IDNO.:9) fused directly to Met 1 of PRMT8.

Protein Expression.

E. coli (BL21(DE3) Gold, Stratagene) made competent by the CaCl₂ methodwere transformed with the PRMT8 construct and ampicillin selection.Protein over-expression was accomplished by growing the PRMT8 expressingE. coli clone and inducing expression with 0.3 mM IPTG at 16° C. Theculture was grown for 12 hours, harvested by centrifugation, and storedat −80° C. for purification.

Protein Purification.

Expressed full-length human GST-tagged PRMT8 protein was purified fromcell paste by glutathione sepharose affinity chromatography after theresin was equilibrated with 50 mM phosphate buffer, 200 mM NaCl, 5%glycerol, 5 mM (3-mercaptoethanol, pH7.8 (Buffer A). GST-tagged PRMT8was eluted with 50 mM Tris, 2 mM glutathione, pH 7.8. Pooled fractionswere cleaved by thrombin (10 U) and dialysed in buffer A. GST wasremoved by reloading the cleaved protein sample onto glutathionesepharose column and PRMT8 was collected in the flow-through fractions.PRMT8 was purified further by ceramic hydroxyapatite chromatography. Thecolumn was washed with 50 mM phosphate buffer, 100 mM NaCl, 5% glycerol,5 mM β-mercaptoethanol, pH 7.8 and PRMT8 was eluted by 100 mM phosphatein the same buffer. Protein was concentrated and buffer was exchanged to50 mM Tris, 300 mM NaCl, 10% glycerol, 5 mM 3-mercaptoethanol, pH 7.8 byultrafiltration. The purity of recovered protein was 89%.

Predicted Translations:

GST-tagged PRMT8 (SEQ ID NO.: 10)MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLVPRGSPEFMGMKHSSRCLLLRRKMAENAAESTEVNSPPSQPPQPVVPAKPVQCVHHVSTQPSCPGRGKMSKLLNPEEMTSRDYYFDSYAHFGIHEEMLKDEVRTLTYRNSMYHNKHVFKDKVVLDVGSGTGILSMFAAKAGAKKVFGIECSSISDYSEKIIKANHLDNIITIFKGKVEEVELPVEKVDIIISEWMGYCLFYESMLNTVIFARDKWLKPGGLMFPDRAALYVVAIEDRQYKDFKIHWWENVYGFDMTCIRDVAMKEPLVDIVDPKQVVTNACLIKEVDIYTVKTEELSFTSAFCLQIQRNDYVHALVTYFNIEFTKCHKKMGFSTAPDAPYTHWKQTVFYLEDYLTVRRGEEIYGTISMKPNAKNVRDLDFTVDLDFKGQLCETSVSNDYKMR

General Procedure for PRMT8 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween 20, prepared on theday of use. Compounds in 100% DMSO (1 ul) were spotted into apolypropylene 384-well V-bottom plates (Greiner) using a Platemate Plusoutfitted with a 384-channel head (Thermo Scientific). DMSO (1 ul) wasadded to Columns 11, 12, 23, 24, rows A-H for the maximum signal controland 1 ul of SAH, a known product and inhibitor of PRMT8, was added tocolumns 11, 12, 23, 24, rows I-P for the minimum signal control. Acocktail (40 ul) containing the PRMT8 enzyme was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate with PRMT8for 30 min at room temperature, then a cocktail (10 ul) containing³H-SAM and peptide was added to initiate the reaction (final volume=51ul). The final concentrations of the components were as follows: PRMT8was 1.5 nM, ³H-SAM was 50 nM, non-radiolabeled SAM was 550 nM, peptidewas 150 nM, SAH in the minimum signal control wells was 1 mM, and theDMSO concentration was 2%. The assays were stopped by the addition ofnon-radiolabeled SAM (10 ul) to a final concentration of 400 uM, whichdilutes the ³H-SAM to a level where its incorporation into the peptidesubstrate is no longer detectable. 50 ul of the reaction in the 384-wellpolypropylene plate was then transferred to a 384-well Flashplate andthe biotinylated peptides were allowed to bind to the streptavidinsurface for at least 1 hour before being washed once with 0.1% Tween20in a Biotek ELx405 plate washer. The plates were then read in aPerkinElmer TopCount plate reader to measure the quantity of ³H-labeledpeptide bound to the Flashplate surface, measured as disintegrations perminute (dpm) or alternatively, referred to as counts per minute (cpm).

% Inhibition Calculation

${\% \mspace{14mu} {inh}} = {100 - {( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} ) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{( {{Top} - {Bottom}} )}{( {1 + ( \frac{X}{{IC}_{50}} )^{{Hill}\mspace{14mu} {Coefficient}}} }}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

PRMT3 Biochemical Assay

General Materials.

S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), bicine,Tween20, dimethylsulfoxide (DMSO), bovine skin gelatin (BSG),),isopropyl-β-D-thiogalactopyranoside (IPTG), andTris(2-carboxyethyl)phosphine hydrochloride solution (TCEP) werepurchased from Sigma-Aldrich at the highest level of purity possible.³H-SAM was purchase from American Radiolabeled Chemicals with a specificactivity of 80 Ci/mmol. 384-well streptavidin Flashplates were purchasedfrom PerkinElmer.

Substrates.

Peptide containing the classic RMT substrate motif was synthesized withan N-terminal linker-affinity tag motif and a C-terminal amide cap by21^(st) Century Biochemicals. The peptide was purified byhigh-performance liquid chromatography (HPLC) to greater than 95% purityand confirmed by liquid chromatography mass spectrometry (LC-MS). Thesequence was Biot-Ahx-GGRGGFGGRGGFGGRGGFG-amide (SEQ ID NO.:11).

Molecular Biology:

Full-length human PRMT3 (NM_005788.3) isoform 1 transcript clone wasamplified from an HEK 293 cDNA library and subcloned into pGEX-KG (GELife Sciences). The resulting construct encodes an N-terminal GST tagand a thrombin cleavage sequence(MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLVPRG S) (SEQ IDNO.:12) fused directly to Cys 2 of PRMT3.

Protein Expression.

E. coli (BL21(DE3) Gold, Stratagene) made competent by the CaCl₂ methodwere transformed with the PRMT3 construct and ampicillin selection.Protein over-expression was accomplished by growing the PRMT3 expressingE. coli clone and inducing expression with 0.3 mM IPTG at 16° C. Theculture was grown for 12 hours, harvested by centrifugation, and storedat −80° C. for purification.

Protein Purification.

Expressed full-length human GST-tagged PRMT3 protein was purified fromcell paste by glutathione sepharose affinity chromatography afterequilibration of the resin with 50 mM phosphate buffer, 200 mM NaCl, 5%glycerol, 1 mM EDTA, 5 mM β-mercaptoethanol, pH6.5 (Buffer A).GST-tagged PRMT3 was eluted with 50 mM Tris, 2 mM glutathione, pH 7.1and 50 mM Tris, 20 mM glutathione, pH 7.1. Pooled fractions weredialysed in 20 mM Tris, 50 mM NaCl, 5% glycerol, 1 mM EDTA, 1 mM DTT,pH7.5 (Buffer B) and applied to a Q Sepharose Fast Flow column.GST-tagged PRMT3 was eluted by 500 mM NaCl in buffer B. Pooled fractionswere dialyzed in 25 mM phosphate buffer, 100 mM NaCl, 5% glycerol, 2 mMDTT, pH 6.8 (Buffer C) and loaded on to a ceramic hydroxyapatite column.GST-tagged PRMT3 eluted with 25-400 mM phosphate in buffer C. Proteinwas concentrated and buffer was exchanged to 20 mM Tris, 150 mM NaCl, 5%glycerol, 5 mM (3-mercaptoethanol, pH7.8 by ultrafiltration. The purityof recovered protein was 70%.

Predicted Translations:

GST-tagged PRMT3 (SEQ ID NO.: 13)MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLVPRGSCSLASGATGGRGAVENEEDLPELSDSGDEAAWEDEDDADLPHGKQQTPCLFCNRLFTSAEETFSHCKSEHQFNIDSMVHKHGLEFYGYIKLINFIRLKNPTVEYMNSIYNPVPWEKEEYLKPVLEDDLLLQFDVEDLYEPVSVPFSYPNGLSENTSVVEKLKHMEARALSAEAALARAREDLQKMKQFAQDFVMHTDVRTCSSSTSVIADLQEDEDGVYFSSYGHYGIHEEMLKDKIRTESYRDFIYQNPHIFKDKVVLDVGCGTGILSMFAAKAGAKKVLGVDQSEILYQAMDIIRLNKLEDTITLIKGKIEEVHLPVEKVDVIISEWMGYFLLFESMLDSVLYAKNKYLAKGGSVYPDICTISLVAVSDVNKHADRIAFWDDVYGFKMSCMKKAVIPEAVVEVLDPKTLISEPCGIKHIDCHTTSISDLEFSSDFTLKITRTSMCTAIAGYFDIYFEKNCHNRVVFSTGPQSTKTHWKQTVFLLEKPFSVKAGEALKGKVTVHKNKKDPRSLTVTLTLNNST QTYGLQ

General Procedure for PRMT3 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween 20, prepared on theday of use. Compounds in 100% DMSO (1 ul) were spotted into apolypropylene 384-well V-bottom plates (Greiner) using a Platemate Plusoutfitted with a 384-channel head (Thermo Scientific). DMSO (1 ul) wasadded to Columns 11, 12, 23, 24, rows A-H for the maximum signal controland 1 ul of SAH, a known product and inhibitor of PRMT3, was added tocolumns 11, 12, 23, 24, rows I-P for the minimum signal control. Acocktail (40 ul) containing the PRMT3 enzyme was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate with PRMT3for 30 min at room temperature, then a cocktail (10 ul) containing SAMand peptide was added to initiate the reaction (final volume=51 ul). Thefinal concentrations of the components were as follows: PRMT3 was 0.5nM, ³H-SAM was 100 nM, non-radiolabeled SAM was 1.8 uM, peptide was 330nM, SAH in the minimum signal control wells was 1 mM, and the DMSOconcentration was 2%. The assays were stopped by the addition ofpotassium chloride (10 ul) to a final concentration of 100 mM. 50 ul ofthe reaction in the 384-well polypropylene plate was then transferred toa 384-well Flashplate and the biotinylated peptides were allowed to bindto the streptavidin surface for at least 1 hour before being washed oncewith 0.1% Tween20 in a Biotek ELx405 plate washer. The plates were thenread in a PerkinElmer TopCount plate reader to measure the quantity of³H-labeled peptide bound to the Flashplate surface, measured asdisintegrations per minute (dpm) or alternatively, referred to as countsper minute (cpm).

% Inhibition Calculation

${\% \mspace{14mu} {inh}} = {100 - {( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} ) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{( {{Top} - {Bottom}} )}{( {1 + ( \frac{X}{{IC}_{50}} )^{{Hill}\mspace{14mu} {Coefficient}}} }}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

CARM1 Biochemical Assay

General Materials.

S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), bicine,Tween20, dimethylsulfoxide (DMSO), bovine skin gelatin (BSG), sodiumbutyrate and Tris(2-carboxyethyl)phosphine hydrochloride solution (TCEP)were purchased from Sigma-Aldrich at the highest level of puritypossible. ³H-SAM was purchase from American Radiolabeled Chemicals witha specific activity of 80 Ci/mmol. 384-well streptavidin Flashplateswere purchased from PerkinElmer.

Substrates.

Peptide representative of human histone H3 residues 16-30 wassynthesized with an N-terminal linker-affinity tag motif and aC-terminal amide cap by 21^(st) Century Biochemicals. The peptide waspurified by high-performance liquid chromatography (HPLC) to greaterthan 95% purity and confirmed by liquid chromatography mass spectrometry(LC-MS). The sequence was Biot-Ahx-PRKQLATKAARKSAP-amide and contained amonomethylated arginine at position 26 (SEQ ID NO.:14).

Molecular Biology:

Human CARM1 (PRMT4) (NM_199141.1) transcript clone was amplified from anHEK 293 cDNA library, incorporating a flanking 5′ sequence encoding aFLAG tag (MDYKDDDDK) (SEQ ID NO.:6) fused directly to Ala 2 of CARM1 and3′ sequence encoding a hexa His sequence (EGHHHHHH) (SEQ ID NO.:15)fused directly to Ser 608. The gene sequence encoding isoform1containing a deletion of amino acids 539-561 was amplified subsequentlyand subcloned into pFastBacMam (Viva Biotech).

Protein Expression.

Recombinant baculovirus were generated according to Bac-to-Bac kitinstructions (Life Technologies). Protein over-expression wasaccomplished by infecting exponentially growing HEK 293F cell culture at1.3×10⁶ cell/ml with virus (MOI=10) in the presence of 8 mM sodiumbutyrate. Infections were carried out at 37° C. for 48 hours, harvestedby centrifugation, and stored at −80° C. for purification.

Protein Purification.

Expressed full-length human Flag- and His-tagged CARM1 protein waspurified from cell paste by anti-flag M2 affinity chromatography withresin equilibrated with buffer containing 20 mM Tris, 150 mM NaCl, 5%glycerol, pH 7.8. Column was washed with 500 mM NaCl in buffer A andFlag-CARM1-His was eluted with 200 ug/ml FLAG peptide in buffer A.Pooled fractions were dialyzed in 20 mM Tris, 150 mM NaCl, 5% glyceroland 1 mM DTT, pH 7.8. The purity of recovered protein was 94.

Predicted Translations:

Flag-CARM1-His (SEQ ID NO.: 16)MDYKDDDDKAAAAAAVGPGAGGAGSAVPGGAGPCATVSVFPGARLLTIGDANGEIQRHAEQQALRLEVRAGPDSAGIALYSHEDVCVFKCSVSRETECSRVGKQSFIITLGCNSVLIQFATPNDFCSFYNILKTCRGHTLERSVFSERTEESSAVQYFQFYGYLSQQQNMMQDYVRTGTYQRAILQNHTDFKDKIVLDVGCGSGILSFFAAQAGARKIYAVEASTMAQHAEVLVKSNNLTDRIVVIPGKVEEVSLPEQVDIIISEPMGYMLFNERMLESYLHAKKYLKPSGNMFPTIGDVHLAPFTDEQLYMEQFTKANFWYQPSFHGVDLSALRGAAVDEYFRQPVVDTFDIRILMAKSVKYTVNFLEAKEGDLHRIEIPFKFHMLHSGLVHGLAFWFDVAFIGSIMTVWLSTAPTEPLTHWYQVRCLFQSPLFAKAGDTLSGTCLLIANKRQSYDISIVAQVDQTGSKSSNLLDLKNPFFRYTGTTPSPPPGSHYTSPSENMWNTGSTYNLSSGMAVAGMPTAYDLSSVIASGSSVGHNNLIPLGSSGAQGSGGGSTSAHYAVNSQFTMGGPAISMASPMSIPTNTMHYGSEGHHHHH H

General Procedure for CARM1 Enzyme Assays on Peptide Substrates.

The assays were all performed in a buffer consisting of 20 mM Bicine(pH=7.6), 1 mM TCEP, 0.005% BSG, and 0.002% Tween 20, prepared on theday of use. Compounds in 100% DMSO (1 ul) were spotted into apolypropylene 384-well V-bottom plates (Greiner) using a Platemate Plusoutfitted with a 384-channel head (Thermo Scientific). DMSO (1 ul) wasadded to Columns 11, 12, 23, 24, rows A-H for the maximum signal controland 1 ul of SAH, a known product and inhibitor of CARM1, was added tocolumns 11, 12, 23, 24, rows I-P for the minimum signal control. Acocktail (40 ul) containing the CARM1 enzyme was added by MultidropCombi (Thermo-Fisher). The compounds were allowed to incubate with CARM1for 30 min at room temperature, then a cocktail (10 ul) containing³H-SAM and peptide was added to initiate the reaction (final volume=51ul). The final concentrations of the components were as follows: CARM1was 0.25 nM, ³H-SAM was 30 nM, peptide was 250 nM, SAH in the minimumsignal control wells was 1 mM, and the DMSO concentration was 2%. Theassays were stopped by the addition of non-radiolabeled SAM (10 ul) to afinal concentration of 300 uM, which dilutes the ³H-SAM to a level whereits incorporation into the peptide substrate is no longer detectable. 50ul of the reaction in the 384-well polypropylene plate was thentransferred to a 384-well Flashplate and the biotinylated peptides wereallowed to bind to the streptavidin surface for at least 1 hour beforebeing washed once with 0.1% Tween20 in a Biotek ELx405 plate washer. Theplates were then read in a PerkinElmer TopCount plate reader to measurethe quantity of ³H-labeled peptide bound to the Flashplate surface,measured as disintegrations per minute (dpm) or alternatively, referredto as counts per minute (cpm).

% Inhibition Calculation

${\% \mspace{14mu} {inh}} = {100 - {( \frac{{dpm}_{cmpd} - {dpm}_{\min}}{{dpm}_{\max} - {dpm}_{\min}} ) \times 100}}$

Where dpm=disintegrations per minute, cmpd=signal in assay well, and minand max are the respective minimum and maximum signal controls.

Four-Parameter IC50 Fit

$Y = {{Bottom} + \frac{( {{Top} - {Bottom}} )}{( {1 + ( \frac{X}{{IC}_{50}} )^{{Hill}\mspace{14mu} {Coefficient}}} }}$

Where top and bottom are the normally allowed to float, but may be fixedat 100 or 0 respectively in a 3-parameter fit. The Hill Coefficientnormally allowed to float but may also be fixed at 1 in a 3-parameterfit. Y is the % inhibition and X is the compound concentration.

TABLE 2 Biochemical IC₅₀ Cmpd No. PRMT1 PRMT6 PRMT8 PRMT3 CARM1 1 A A BD B 2 A A B C B 3 — A B D C 4 A A B E B 5 A A B C B 6 A A B D B 7 A A BD A 8 A A B D B 9 A A B D C 10 B B C E E 11 A A B C A 12 C C E E E 13 AA B D B 14 A B B D B 15 A A B C B 16 A B B D B 17 A A B C A 18 A A A C A19 B B C D D 20 A A B C B 21 C B D E E 22 A A B D D 23 A A B D C 24 A BB E — 25 B B C — — 26 — B D — — 27 — C E — — 28 B A C — — 29 B B C — —30 — B D — — 31 — D E — — 32 B A C — — 33 A A B — — 34 A A B — — 35 A AB — — 36 A A B — — 37 A A B — — 38 A A B — — 39 — A B — — 40 — A B — —41 A A D — — 42 A A B — — 43 — A — — — 44 C B — — — 45 A A — — — 46 A A— — — 47 A A — — — 48 A A — — — 49 A A — — — 50 A A — — — 51 A A — — —52 A A — — — 53 A A — — — 54 A A — — — 55 A A — — — 56 A A — — — 57 A A— — — 58 A A — — — 59 A B — — — 60 A B — — — 61 A A — — — 62 A A A — —63 A A A — — 64 — — — — — 65 A A A — — 66 — — — — — 67 A A A — — 68 — —— — — 69 — — — — — 70 A A A — — 71 — — — — — 72 — — — — — 73 — — — — —74 — — — — — 75 — — — — — 76 — — — — — 77 — — — — — 78 — — — — — 79 — —— — — 80 — — — — — 81 — — — — — 82 — — — — — 83 — — — — — 84 — — — — —85 — — — — — 86 C D D — — 87 B D C — — 88 — — — — — 89 — — — — — 90 A BB — — 91 A A A — — 92 A A A — — 93 A A A — — 94 — — — — — 95 A A A — —96 — — — — — 97 — — — — — 98 — — — — — 99 — — — — — 100 — — — — — 101 —— — — — 102 — — — — — 103 — — — — — 104 A A A — — 105 — — — — — 106 A AA — — 107 A B A — — 108 A B B — — 109 A B A — — 110 A A A — — 111 A A A— — 112 A A A — — 113 A A B — — 114 B A B — — 115 A A B — — 116 A A A —— 117 A A B — — 118 C E E — — 119 B C B — — 120 A A A — — 121 A A A — —122 B E D — — 123 A A A — — 124 A A A — — 125 A A A — — 126 A A A — —127 A A A — — 128 A A A — — 129 A A A — — 130 A A A — — 131 B E C — —132 B D D — — 133 A A A — — 134 A A A — — 135 A A A — — 136 A A A — —137 A A A — — 138 B C C — — 139 A A B — — 140 A A A — — 141 A A A — —142 A A A — — 143 A A A — — 144 A A A — — 145 A A — — — 146 A A — — —147 A A — — — 148 A A — — — 149 A A — — — 150 A A — — — 151 A A — — —152 A A — — — 153 A A — — — 154 A A — — — 155 A A — — — 156 A A — — —157 A A — — — 158 A A — — — 159 A A — — — 160 A B — — — 161 A A A — —162 A A A — — 163 A A A — — 164 A A A — — 165 A B A — — 166 A A A — —167 A A A — — 168 A A A — — 169 A A A — — 170 A A A — — 171 A A — — —172 A A — — — 173 A A — — — 174 A A — — — 175 A A A — — 176 A A A — —177 A B B — — 178 A A A — — 179 A A A — — 180 A A — — — 181 A A — — —182 A A A — — 183 A A A — — 184 A B A — — 185 A A A — — 186 B B A — —187 B B B — — 188 B C C — — 189 B C B — — 190 B B B — — 191 A B B — —192 B B B — — 193 A A A — — 194 A A A — — 195 A A A — — 196 B B B — —197 A A A — — 198 D D D — — 199 A A A — — 200 A A A — — 201 A B A — —202 A A A — — 203 A A A — — 204 A A A — — 205 A A A — — 206 A B A — —207 A A A — — 208 A A A — — 209 A A A — — 210 A A A — — 211 A A A — —212 A A A — — 213 A A B — — 214 A A A — — 215 A A A — — 216 A A A — —217 A A A — — 218 A A A — — 219 A A A — — 220 A A A — — 221 A A A — —222 A A A — — 223 A A A — — 224 A A A — — 225 A A A — — 226 A A A — —227 A A A — — 228 A A A — — 229 A A A — — 230 A A A — — 231 A A A — —232 A A A — — 233 A A A — — 234 A A A — — 235 A A A — — 236 A A A — —237 A A A — — 238 A A A — — 239 A A A — — 240 A A A — — 241 A A A — —242 A A A — — 243 A A A — — 244 A A B — — 245 A A A — — 246 A A A — —247 A A A — — 248 A A A — — 249 A A A — — 250 A A A — — 251 A A A — —252 A A A — — 253 A A A — — 254 A A A — — 255 A A A — — 256 A A A — —257 A B A — — 258 A A A — — 259 A A A — — 260 A A A — — 261 A A A — —262 A A A — — 263 A A A — — 264 A B B — — 265 A B B — — 266 A A A — —267 D D D — — 268 A B B — — 269 A B B — — 270 A B B — — 271 A B A — —272 A A A — — 273 A A A — — 274 A B A — — 275 A A A — — “—” indicates nodata provided. For Table 2, “A” indicates an IC₅₀ ≤ 0.100 μM, “B”indicates an IC₅₀ of 0.101-1.00 μM, “C” indicates an IC₅₀ of 1.01-3.00μM, “D” indicates an IC₅₀ of 3.01-10 μM, and IC₅₀ ≥ 10.01 μM.

TABLE 2a Biochemical IC₅₀ (Numerical, μM)* Cmpd No. PRMT1 PRMT6 PRMT8PRMT3 CARM1 1 0.01 0.07 0.39 6.83 0.26 2 0.01 0.04 0.17 3.00 0.11 3 —0.02 0.95 9.40 1.23 4 0.02 0.02 0.59 >10 0.50 5 0.01 0.00 0.42 1.45 0.316 0.04 0.03 0.24 3.94 0.57 7 0.01 0.02 0.52 5.91 0.09 8 0.02 0.04 0.365.81 0.37 9 0.01 0.01 0.76 8.93 3.00 10 0.39 0.31 1.92 >10 >10 11 0.010.05 0.25 1.58 0.05 12 1.58 1.08 >10 >10 >10 13 0.01 0.01 0.30 3.62 0.2014 0.09 0.93 0.66 6.77 0.51 15 0.05 0.04 0.70 2.34 0.31 16 0.02 0.150.69 5.79 0.11 17 0.01 0.05 0.41 2.36 0.09 18 0.00 0.01 0.04 1.75 0.1019 0.19 0.17 2.30 7.74 3.78 20 0.02 0.02 0.47 2.34 0.25 21 1.49 0.679.56 >10 >10 22 0.01 0.02 0.30 6.46 3.77 23 0.01 0.01 0.28 9.65 2.78 240.02 0.18 0.30 >10 — 25 0.25 0.29 2.86 — — 26 — 0.51 5.63 — — 27 —2.41 >10 — — 28 0.18 0.08 1.68 — — 29 — 0.29 3.81 — — 30 — 6.59 >10 — —31 0.11 0.03 1.15 — — 32 0.02 0.02 0.77 — — 33 0.02 0.02 0.50 — — 340.02 0.03 0.44 — — 35 0.00 0.01 0.14 — — 36 0.00 0.01 0.28 — — 37 0.010.01 0.13 — — 38 0.00 0.01 0.26 — — 39 — 0.01 0.36 — — 40 0.02 0.02 7.01— — 41 0.01 0.00 0.35 — — 42 — 0.02 0.0300 — — 43 — 0.00 0.0116 — — 44 —0.02 0.0624 — — 45 0.0028 0.0056 0.0053 — — 46 0.0079 0.0122 0.0062 — —47 0.0043 0.0173 0.0125 — — 48 0.0074 0.0178 0.0083 — — 49 0.0043 0.01990.0073 — — 50 0.0086 0.0206 0.0112 — — 51 0.0094 0.0239 0.0214 — — 520.0047 0.0243 0.0104 — — 53 0.0105 0.0380 0.0128 — — 54 0.0080 0.03820.0247 — — 55 0.0128 0.0404 0.0247 — — 56 0.0138 0.0618 0.0320 — — 570.0186 0.0694 0.0301 — — 58 0.0265 0.1077 0.0721 — — 59 0.0196 0.12770.0484 — — 60 0.0275 0.1549 0.1286 — — 61 0.0205 0.0491 — — — 62 0.011870.02902 0.02 — — 63 0.0090 0.0121 0.0120 — — 64 — — — — — 65 0.009650.04193 0.01355 — — 66 — — — — — 67 0.0078 0.0180 0.0119 — — 68 — — — —— 69 — — — — — 70 0.0092 0.0440 0.0242 — — 71 — — — — — 72 — — — — — 73— — — — — 74 — — — — — 75 — — — — — 76 — — — — — 77 — — — — — 78 — — — —— 79 — — — — — 80 — — — — — 81 — — — — — 82 — — — — — 83 — — — — — 84 —— — — — 85 — — — — — 86 1.94296 3.40906 3.96737 — — 87 0.56419 3.726331.01384 — — 88 — — — — — 89 — — — — — 90 0.0344 0.12244 0.11687 — — 910.01668 0.06307 0.02896 — — 92 0.0209 0.04904 0.04206 — — 93 0.031280.07193 0.08812 — — 94 — — — — — 95 0.00935 0.0692 0.01887 — — 96 — — —— — 97 — — — — — 98 — — — — — 99 — — — — — 100 — — — — — 101 — — — — —102 — — — — — 103 — — — — — 104 0.00804 0.04046 0.01158 — — 105 — — — —— 106 0.0095 0.01348 0.0145 — — 107 0.0212 0.1296 0.0554 — — 108 0.06100.3255 0.2521 — — 109 0.0267 0.1243 0.0560 — — 110 0.0131 0.0294 0.0429— — 111 0.0224 0.0455 0.0599 — — 112 0.0189 0.0574 0.0692 — — 113 0.05720.0669 0.1193 — — 114 0.1009 0.0926 0.3400 — — 115 0.0887 0.0627 0.1964— — 116 0.0098 0.0187 0.0118 — — 117 0.0386 0.0962 0.1428 — — 1182.1552 >10.0 >10.0 — — 119 0.1239 2.7522 0.7278 — — 120 0.0101 0.04280.0251 — — 121 0.0152 0.0977 0.0326 — — 122 0.4207 >10.0 8.3743 — — 1230.0088 0.0180 0.0247 — — 124 0.0064 0.0262 0.0108 — — 125 0.0053 0.01310.0090 — — 126 0.0040 0.0094 0.0065 — — 127 0.0165 0.0223 0.0478 — — 1280.0052 0.0131 0.0082 — — 129 0.0236 0.0297 0.0837 — — 130 0.0034 0.00420.0045 — — 131 0.2699 >10.0 1.9689 — — 132 0.6264 5.4954 4.3316 — — 1330.0059 0.0124 0.0091 — — 134 0.0139 0.0351 0.0248 — — 135 0.0121 0.03490.0317 — — 136 0.0029 0.0072 0.0068 — — 137 0.0073 0.0233 0.0162 — — 1380.3486 1.2075 1.4305 — — 139 0.0305 0.0226 0.1157 — — 140 0.0156 0.02400.0339 — — 141 0.0107 0.0230 0.0297 — — 142 0.0072 0.0199 0.0159 — — 1430.0106 0.0220 0.0330 — — 144 0.0216 0.0356 0.0784 — — 145 0.0262 0.0380— — — 146 0.0140 0.0217 — — — 147 0.0183 0.0327 — — — 148 0.0048 0.0081— — — 149 0.0045 0.0072 — — — 150 0.0440 0.0556 — — — 151 0.0369 0.0477— — — 152 0.0142 0.0352 — — — 153 0.0232 0.0284 — — — 154 0.0177 0.0443— — — 155 0.0315 0.0608 — — — 156 0.0172 0.0578 — — — 157 0.0185 0.0479— — — 158 0.0202 0.0910 — — — 159 0.0197 0.0538 — — — 160 0.0218 0.1121— — — 161 0.0256 0.0729 0.0374 — — 162 0.0389 0.0610 0.0927 — — 1630.0173 0.0657 0.0158 — — 164 0.0197 0.0708 0.0191 — — 165 0.0176 0.12040.0181 — — 166 0.0180 0.0486 0.0230 — — 167 0.0110 0.0315 0.0130 — — 1680.0198 0.0451 0.0281 — — 169 0.0065 0.0178 0.0102 — — 170 0.0210 0.05780.0237 — — 171 0.0147 0.0411 — — — 172 0.0230 0.0458 — — — 173 0.02320.0899 — — — 174 0.0155 0.0844 — — — 175 0.0133 0.0376 0.0296 — — 1760.0102 0.0364 0.0286 — — 177 0.0276 0.1129 0.1499 — — 178 0.0058 0.01570.0205 — — 179 0.0159 0.0629 0.0573 — — 180 0.0049 0.0147 — — — 1810.0047 0.0279 — — — 182 0.01161 0.0735 0.03227 — — 183 0.01348 0.087680.03943 — — 184 0.01977 0.10057 0.04732 — — 185 0.01421 0.03518 0.02442— — 186 0.11047 0.10652 0.08958 — — 187 0.19316 0.26079 0.18533 — — 1880.27493 1.23016 1.53507 — — 189 0.10809 1.47313 0.47825 — — 190 0.123940.15596 0.35151 — — 191 0.0504 0.10664 0.15195 — — 192 0.21645 0.172680.45873 — — 193 0.0284 0.09512 0.06882 — — 194 0.01446 0.02273 0.03525 —— 195 0.01444 0.05169 0.0356 — — 196 0.14877 0.11139 0.41502 — — 1970.00827 0.01065 0.01563 — — 198 10 8.53693 10 — — 199 0.01683 0.036530.03833 — — 200 0.01152 0.04163 0.01623 — — 201 0.02286 0.13436 0.03183— — 202 0.01266 0.06637 0.02686 — — 203 0.02837 0.03683 0.08436 — — 2040.01837 0.06361 0.05164 — — 205 0.01283 0.07151 0.01537 — — 206 0.019460.2066 0.02894 — — 207 0.00568 0.01846 0.00913 — — 208 0.00694 0.045270.01276 — — 209 0.02776 0.05208 0.04385 — — 210 0.01313 0.03166 0.02286— — 211 0.00828 0.01138 0.01002 — — 212 0.00557 0.01357 0.00828 — — 2130.05388 0.09132 0.16503 — — 214 0.01978 0.05049 0.02743 — — 215 0.014060.0306 0.02297 — — 216 0.01208 0.0205 0.01779 — — 217 0.03018 0.034040.05952 — — 218 0.0417 0.03035 0.06702 — — 219 0.00896 0.03189 0.01499 —— 220 0.00929 0.01841 0.01294 — — 221 0.00644 0.01915 0.01545 — — 2220.04299 0.02131 0.09002 — — 223 0.00665 0.01575 0.00865 — — 224 0.009270.01805 0.01018 — — 225 0.00762 0.01164 0.01389 — — 226 0.03311 0.041490.05758 — — 227 0.0168 0.01782 0.03576 — — 228 0.069 0.01848 0.10005 — —229 0.01084 0.02912 0.01595 — — 230 0.01346 0.02307 0.02574 — — 2310.00649 0.01474 0.00932 — — 232 0.00911 0.01937 0.02101 — — 233 0.020750.04787 0.04267 — — 234 0.01611 0.08411 0.04276 — — 235 0.02571 0.034080.04404 — — 236 0.02903 0.06239 0.05783 — — 237 0.04219 0.04098 0.08329— — 238 0.0296 0.05387 0.08009 — — 239 0.00655 0.00775 0.01319 — — 2400.0061 0.00672 0.01091 — — 241 0.0224 0.03276 0.04894 — — 242 0.028340.04166 0.08368 — — 243 0.03244 0.01514 0.07082 — — 244 0.03978 0.054820.11313 — — 245 0.01854 0.06216 0.01834 — — 246 0.01258 0.0283 0.02378 —— 247 0.01107 0.02018 0.01644 — — 248 0.01319 0.03262 0.02818 — — 2490.02419 0.08278 0.08453 — — 250 0.02396 0.01435 0.04241 — — 251 0.013050.01711 0.02073 — — 252 0.03086 0.04826 0.05152 — — 253 0.00715 0.013350.01168 — — 254 0.01728 0.01764 0.044 — — 255 0.01016 0.02765 0.02448 —— 256 0.02367 0.03851 0.04405 — — 257 0.02982 0.15318 0.03096 — — 2580.00759 0.01798 0.01083 — — 259 0.01579 0.0331 0.01706 — — 260 0.020210.08314 0.03733 — — 261 0.013 0.03165 0.01668 — — 262 0.03373 0.08120.07236 — — 263 0.01915 0.0638 0.03513 — — 264 0.07798 0.1327 0.18082 —— 265 0.05883 0.17447 0.17058 — — 266 0.0193 0.07899 0.0631 — — 2673.4983 10 5.91407 — — 268 0.06863 0.14146 0.20985 — — 269 0.051270.10664 0.15456 — — 270 0.06247 0.14863 0.20383 — — 271 0.03706 0.12080.10019 — — 272 0.01236 0.07387 0.02578 — — 273 0.01145 0.04045 0.02257— — 274 0.00819 0.10255 0.01193 — — 275 0.00462 0.01859 0.01038 — — *ForTable 2a, numerical values represent data from a single experiment or anaverage of multiple experiments.

RKO Methylation Assay

RKO adherent cells were purchased from ATCC (American Type CultureCollection), Manassas, Va., USA. DMEM/Glutamax medium,penicillin-streptomycin, heat inactivated fetal bovine serum, 0.05%trypsin and D-PBS were purchased from Life Technologies, Grand Island,N.Y., USA. Odyssey blocking buffer, 800CW goat anti-rabbit IgG (H+L)antibody, and Licor Odyssey infrared scanner were purchased from LicorBiosciences, Lincoln, Nebr., USA. Mono-methyl arginine antibody waspurchased from Cell Signaling Technology, Danvers, Mass., USA. Methanolwas purchased from VWR, Franklin, Mass., USA. 10% Tween 20 was purchasedfrom KPL, Inc., Gaithersburg, Md., USA. DRAQ5 was purchased fromBiostatus Limited, Leicestershire, UK.

RKO adherent cells were maintained in growth medium (DMEM/Glutamaxmedium supplemented with 10% v/v heat inactivated fetal bovine serum and100 units/mL penicillin-streptomycin) and cultured at 37° C. under 5%CO₂.

Cell Treatment, in Cell Western (ICW) for Detection of Mono-MethylArginine and DNA Content.

RKO cells were seeded in assay medium at a concentration of 20,000 cellsper mL to a poly-D-lysine coated 384 well culture plate (BD Biosciences356697) with 50 μL per well. Compound (100 nL) from a 96-well sourceplate was added directly to 384 well cell plate. Plates were incubatedat 37° C., 5% CO₂ for 72 hours. After three days of incubation, plateswere brought to room temperature outside of the incubator for tenminutes and blotted on paper towels to remove cell media. 50 μL of icecold 100% methanol was added directly to each well and incubated for 30min at room temperature. After 30 min, plates were transferred to aBiotek EL406 plate washer and washed 2 times with 100 μL per well ofwash buffer (1×PBS). Next 60 μL per well of Odyssey blocking buffer(Odyssey Buffer with 0.1% Tween 20 (v/v)) were added to each plate andincubated 1 hour at room temperature. Blocking buffer was removed and 20μL per well of primary antibody was added (mono-methyl arginine diluted1:200 in Odyssey buffer with 0.1% Tween 20 (v/v)) and plates wereincubated overnight (16 hours) at 4° C. Plates were washed 5 times with100 μL per well of wash buffer. Next 20 μL per well of secondaryantibody was added (1:200 800CW goat anti-rabbit IgG (H+L) antibody,1:1000 DRAQ5 (Biostatus limited) in Odyssey buffer with 0.1% Tween 20(v/v)) and incubated for 1 hour at room temperature. The plates werewashed 5 times with 100 μL per well wash buffer then 2 times with 100 μLper well of water. Plates were allowed to dry at room temperature thenimaged on the Licor Odyssey machine which measures integrated intensityat 700 nm and 800 nm wavelengths. Both 700 and 800 channels werescanned.

Calculations:

First, the ratio for each well was determined by:

$( \frac{{monomethyl}\mspace{14mu} {Arginine}\mspace{14mu} 800\mspace{14mu} {nm}\mspace{14mu} {value}}{{DRAQ}\; 5\mspace{14mu} 700\mspace{14mu} {nm}\mspace{14mu} {value}} )$

Each plate included fourteen control wells of DMSO only treatment(minimum activation) as well as fourteen control wells for maximumactivation treated with 20 μM of a reference compound. The average ofthe ratio values for each control type was calculated and used todetermine the percent activation for each test well in the plate.Reference compound was serially diluted three-fold in DMSO for a totalof nine test concentrations, beginning at 20 μM. Percent activation wasdetermined and EC₃₀ curves were generated using triplicate wells perconcentration of compound.

${{Percent}\mspace{14mu} {Activation}} = {100 - ( {( \frac{( {{Individual}\mspace{14mu} {Test}\mspace{14mu} {Sample}\mspace{14mu} {Ratio}} ) - ( {{Minimum}\mspace{14mu} {Activation}\mspace{14mu} {Ratio}} )}{( {{Maximum}\mspace{14mu} {Activation}\mspace{14mu} {Ratio}} ) - ( {{Minimum}\mspace{14mu} {Activation}\mspace{14mu} {Ratio}} )} )*100} )}$

TABLE 3 In Cell Western Cmpd No. EC₃₀ 9 B 10 C 21 C 22 A 23 A 24 A 25 B26 C 27 C 28 B 29 C 30 C 31 C 32 A 33 B 34 A 35 A 36 A 37 A 38 A 39 A 40A 41 C 42 C 43 A 44 B 45 A 46 A 47 — 48 A 49 — 50 A 51 A 52 — 53 A 54 A55 A 56 — 57 A 58 — 59 — 60 A 61 A 62 A 63 A 64 — 65 A 66 — 67 A 68 — 69— 70 A 71 — 72 — 73 — 74 — 75 — 76 — 77 — 78 — 79 — 80 — 81 — 82 — 83 —84 — 85 — 86 A 87 A 88 — 89 — 90 A 91 A 92 A 93 — 94 — 95 A 96 — 97 — 98— 99 — 100 — 101 — 102 — 103 — 104 A 105 — 106 A 107 — 108 B 109 A 110 A111 A 112 — 113 B 114 — 115 A 116 B 117 — 118 B 119 A 120 A 121 — 122 —123 A 124 A 125 A 126 A 127 A 128 A 129 A 130 — 131 — 132 — 133 A 134 A135 A 136 A 137 A 138 — 139 A 140 A 141 A 142 A 143 A 144 A 145 A 146 A147 B 148 A 149 A 150 A 151 A 152 A 153 A 154 A 155 B 156 A 157 A 158 A159 A 160 A 161 B 162 B 163 A 164 A 165 A 166 A 167 A 168 A 169 A 170 —171 A 172 B 173 A 174 A 175 C 176 A 177 A 178 A 179 A 180 A 181 A 182 A183 A 184 A 185 A 186 — 187 B 188 — 189 B 190 B 191 B 192 B 193 A 194 A195 B 196 B 197 A 198 A 199 A 200 A 201 A 202 A 203 A 204 A 205 A 206 A207 A 208 A 209 A 210 A 211 A 212 A 213 A 214 A 215 A 216 A 217 A 218219 A 220 A 221 A 222 A 223 A 224 A 225 A 226 A 227 A 228 — 229 A 230 A231 A 232 A 233 A 234 A 235 A 236 A 237 A 238 A 239 A 240 A 241 A 242 —243 — 244 A 245 — 246 A 247 A 248 A 249 A 250 — 251 A 252 A 253 A 254 A255 A 256 A 257 A 258 A 259 A 260 A 261 A 262 A 263 — 264 — 265 — 266 —267 — 268 — 269 — 270 — 271 — 272 — 273 — 274 A 275 A “—” indicates nodata provided. For Table 3, “A” indicates an EC₃₀ ≤ 3.00 μM, “B”indicates an EC₃₀ of 3.01-12.00 μM, and “C” indicates an EC₃₀ > 12.01μM.

TABLE 3a In Cell Western (Numerical, μM)* Cmpd No. EC₃₀ 9 5.61 10 >2021 >20 22 1.24 23 1.61 24 2.40 25 11.83 26 >20 27 >20 28 11.10 29 >2030 >20 31 >20 32 2.35 33 3.33 34 1.15 35 0.73 36 0.63 37 0.26 38 0.13 390.50 40 0.87 41 >20 42 >20 43 2.60 44 4.57 45 0.0540 46 0.0080 47 0.052048 0.0140 49 0.0500 50 0.0270 51 0.0520 52 0.0410 53 0.0170 54 0.0870 550.0630 56 0.0960 57 0.0250 58 0.9780 59 0.3570 60 2.8410 61 0.1490 620.16924 63 1.0852 64 — 65 0.07752 66 — 67 0.0810 68 — 69 — 70 0.0500 71— 72 — 73 — 74 — 75 — 76 — 77 — 78 — 79 — 80 — 81 — 82 — 83 — 84 — 85 —86 0.0168 87 2.5254 88 — 89 — 90 0.3158 91 2.05655 92 2.17333 93 — 94 —95 0.12166 96 — 97 — 98 — 99 — 100 — 101 — 102 — 103 — 104 0.01166 105 —106 0.0227 107 — 108 8.5466 109 2.4785 110 2.5472 111 0.9130 112 — 1137.3455 114 — 115 0.0189 116 8.8958 117 — 118 9.5749 119 0.0773 1200.0098 121 — 122 0.3455 123 0.0440 124 0.0214 125 0.0876 126 0.6376 1270.0292 128 1.0750 129 0.0045 130 — 131 — 132 0.0038 133 0.0438 1340.0332 135 1.0140 136 0.0350 137 0.0150 138 — 139 0.6270 140 0.1640 1410.1320 142 0.0520 143 0.0590 144 0.0630 145 2.3000 146 0.1600 147 8.6000148 0.0160 149 0.1060 150 0.2560 151 0.7050 152 0.1190 153 0.1290 1541.7890 155 4.0850 156 1.3570 157 2.4890 158 0.0080 159 0.0630 160 0.1660161 5.0200 162 6.6800 163 0.0660 164 0.0780 165 0.5100 166 0.0830 1670.0180 168 0.0850 169 0.0160 170 — 171 0.1920 172 4.4570 173 0.4080 1740.0850 175 >20 176 2.1420 177 0.5710 178 0.0760 179 0.4760 180 0.0190181 0.1070 182 0.41111 183 0.31865 184 0.45829 185 0.45302 186 — 1873.1603 188 — 189 6.62284 190 6.6228 191 9.24747 192 9.2475 193 1.43822194 1.1848 195 4.88623 196 4.8862 197 0.40624 198 0.4062 199 0.80245 2000.02034 201 0.15455 202 0.32701 203 0.3270 204 1.55419 205 0.01445 2060.07648 207 0.01212 208 0.06719 209 0.91597 210 0.27952 211 0.15488 2120.43283 213 0.4328 214 0.09582 215 0.16359 216 0.11104 217 0.1110 218 —219 0.29433 220 0.2188 221 0.4903 222 2.60878 223 0.27538 224 0.08472225 0.64935 226 1.53074 227 1.5307 228 — 229 0.68908 230 1.31527 2310.09668 232 0.95704 233 1.05613 234 1.0561 235 0.81272 236 0.8127 2371.43678 238 2.15067 239 2.1507 240 1.17758 241 1.1776 242 — 243 — 2442.52536 245 — 246 0.0799 247 0.6979 248 1.3093 249 0.6585 250 — 2511.4511 252 0.2673 253 0.0566 254 1.33631 255 0.21397 256 0.31583 2570.10322 258 0.30272 259 0.40841 260 0.20696 261 0.28926 262 0.2893 263 —264 — 265 — 266 — 267 — 268 — 269 — 270 — 271 — 272 — 273 — 274 0.01681275 0.084 *For Table 3a, numerical values represent data from a singleexperiment or an average of multiple experiments.

Other Embodiments

The foregoing has been a description of certain non-limiting embodimentsof the invention. Those of ordinary skill in the art will appreciatethat various changes and modifications to this description may be madewithout departing from the spirit or scope of the present invention, asdefined in the following claims.

1.-11. (canceled)
 12. A compound of Formula (VI):

or a pharmaceutically acceptable salt thereof, wherein X is N, Z is NR⁴,and Y is CR⁵; or X is NR⁴, Z is N, and Y is CR⁵; or X is CR⁵, Z is NR⁴,and Y is N; or X is CR⁵, Z is N, and Y is NR⁴; R^(x) is optionallysubstituted C₁₋₄ alkyl or optionally substituted C₃₋₄ cycloalkyl; L₁ isa bond, —O—, —N(R^(B))—, —S—, —C(O)—, —C(O)O—, —C(O)S—, —C(O)N(R^(B))—,—C(O)N(R^(B))N(R^(B))—, —OC(O)—, —OC(O)N(R^(B))—, —NR^(B)C(O)—,—NR^(B)C(O)N(R^(B))—, —NR^(B)C(O)N(R^(B))N(R^(B))—, —NR^(B)C(O)O—,—SC(O)—, —C(═NR^(B))—, —C(═NNR^(B))—, —C(═NOR^(A))—,—C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—, —C(S)—, —C(S)N(R^(B))—,—NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—, —N(R^(B))SO₂—,—SO₂N(R^(B))—, or an optionally substituted C₁₋₆ saturated orunsaturated hydrocarbon chain, wherein one or more methylene units ofthe hydrocarbon chain is optionally and independently replaced with —O—,—N(R^(B))—, —S—, —C(O)—, —C(O)O—, —C(O)S—, —C(O)N(R^(B))—,—C(O)N(R^(B))N(R^(B))—, —OC(O)—, —OC(O)N(R^(B))—, —NR^(B)C(O)—,—NR^(B)C(O)N(R^(B))—, —NR^(B)C(O)N(R^(B))N(R^(B))—, —NR^(B)C(O)O—,—SC(O)—, —C(═NR^(B))—, —C(═NNR^(B))—, —C(═NOR^(A))—,—C(═NR^(B))N(R^(B))—, —NR^(B)C(═NR^(B))—, —C(S)—, —C(S)N(R^(B))—,—NR^(B)C(S)—, —S(O)—, —OS(O)₂—, —S(O)₂O—, —SO₂—, —N(R^(B))SO₂—, or—SO₂N(R^(B))—; each R^(A) is independently selected from the groupconsisting of hydrogen, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, optionally substituted heteroaryl, an oxygenprotecting group when attached to an oxygen atom, and a sulfurprotecting group when attached to a sulfur atom; each R^(B) isindependently selected from the group consisting of hydrogen, optionallysubstituted alkyl, optionally substituted alkenyl, optionallysubstituted alkynyl, optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, optionallysubstituted heteroaryl, and a nitrogen protecting group, or an R^(B) andR^(W) on the same nitrogen atom may be taken together with theintervening nitrogen to form an optionally substituted heterocyclicring; Ring A is optionally substituted carbocyclyl, optionallysubstituted heterocyclyl, optionally substituted aryl, or optionallysubstituted heteroaryl; R³ is hydrogen, C₁₋₄ alkyl, or C₃₋₄ cycloalkyl;R⁴ is hydrogen, optionally substituted C₁₋₆ alkyl, optionallysubstituted C₂₋₆ alkenyl, optionally substituted C₂₋₆ alkynyl,optionally substituted C₃₋₇ cycloalkyl, optionally substituted 4-to7-membered heterocyclyl; or optionally substituted C₁₋₄ alkyl-Cy; Cy isoptionally substituted C₃₋₇ cycloalkyl, optionally substituted 4-to7-membered heterocyclyl, optionally substituted aryl, or optionallysubstituted heteroaryl; and R⁵ is hydrogen, halo, —CN, optionallysubstituted C₁₋₄ alkyl, or optionally substituted C₃₋₄ cycloalkyl;wherein, and unless otherwise specified, heterocyclyl or heterocyclicrefers to a radical of a 3-10 membered non-aromatic ring system havingring carbon atoms and 1-4 ring heteroatoms, wherein each heteroatom isindependently selected from nitrogen, oxygen, and sulfur; carbocyclyl orcarbocyclic refers to a radical of a non-aromatic cyclic hydrocarbongroup having from 3 to 10 ring carbon atoms and zero heteroatoms in thenon-aromatic ring system; aryl refers to a radical of a monocyclic orpolycyclic aromatic ring system having 6-14 ring carbon atoms and zeroheteroatoms provided in the aromatic ring system; and heteroaryl refersto a radical of a 5-10 membered monocyclic or bicyclic 4n+2 aromaticring system having ring carbon atoms and 1-4 ring heteroatoms providedin the aromatic ring system, wherein each heteroatom is independentlyselected from nitrogen, oxygen and sulfur.
 13. The compound of claim 12,or a pharmaceutically acceptable salt thereof, wherein the compound isof Formula (VI-a), Formula (VI-b), Formula (VI-c), Formula VI-d),Formula (VI-e), Formula (VI-f), Formula (VI-g), Formula (VI-h), Formula(VI-i), Formula (VI-j), Formula (VI-k), or Formula (VI-l):

or a pharmaceutically acceptable salt thereof; wherein p is 1, 2, 3, 4,5, or
 6. 14.-34. (canceled)
 35. The compound of claim 12, or apharmaceutically acceptable salt thereof, wherein R⁴ is hydrogen oroptionally substituted C₁₋₆ alkyl. 36.-38. (canceled)
 39. The compoundof claim 12, or a pharmaceutically acceptable salt thereof, wherein R⁵is hydrogen or optionally substituted C₁₋₄ alkyl. 40.-51. (canceled) 52.The compound of claim 12, wherein the compound is selected from:

and pharmaceutically acceptable salts thereof.
 53. A pharmaceuticalcomposition comprising a compound of claim 12, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable excipient.54. A kit or packaged pharmaceutical comprising a compound of claim 12,or a pharmaceutically acceptable salt thereof, and instructions for usethereof.
 55. A method of inhibiting an arginine methyl-tranfserase (RMT)comprising contacting a cell with an effective amount of a compound ofclaim 12, or a pharmaceutically acceptable salt thereof. 56.-60.(canceled)
 61. A method of modulating gene expression comprisingcontacting a cell with an effective amount of a compound of claim 12, ora pharmaceutically acceptable salt thereof.
 62. A method of modulatingtranscription comprising contacting a cell with an effective amount of acompound of claim 12, or a pharmaceutically acceptable salt thereof. 63.(canceled)
 64. (canceled)
 65. A method of treating a RMT-mediateddisorder, comprising administering to a subject in need thereof atherapeutically effective amount of a compound of claim 12, or apharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition of claim
 53. 66.-70. (canceled)
 71. The method of claim 65,wherein the disorder is a proliferative disorder, a neurologicaldisorder, a muscular dystrophy, an autoimmune disorder, a vasculardisorder, or a metabolic disorder.
 72. The method of claim 71, whereinthe disorder is cancer or amyotrophic lateral sclerosis. 73.-78.(canceled)
 79. The compound of claim 12, or a pharmaceuticallyacceptable salt thereof, wherein Ring A is of Formula (q-7)-(q-17):

wherein: V²¹, V²², V²³, and V²⁴ are each independently O, S, NR^(N),C═O, or C(R^(C))₂ as valency permits, provided no more than two of V²¹,V²², V²³, and V²⁴ is O, S, or NR^(N), or wherein one of V²¹, V²², V²³,and V²⁴ and another of V²¹, V²², V²³, and V²⁴ adjacent to each other arejoined to form an N═C(R^(C)) or C(R^(C))═C(R^(C)) group provided thering thus formed is not an aromatic ring; each instance of R^(C) isindependently hydrogen, halo, —CN, —NO₂, optionally substituted alkyl,optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted aryl,optionally substituted heterocyclyl, optionally substituted heteroaryl,—OR^(A), —N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; eachinstance of R^(N) is independently hydrogen, optionally substitutedalkyl, optionally substituted alkenyl, optionally substituted alkynyl,optionally substituted carbocyclyl, optionally substituted heterocyclyl,optionally substituted aryl, optionally substituted heteroaryl,—C(═O)R^(A), —C(═O)OR^(A), —C(═O)SR^(A), —C(═O)N(R^(B))₂,—C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —C(═S)R^(A), —C(═S)N(R^(B))₂, —S(═O)R^(A),—SO₂R^(A), —SO₂N(R^(B))₂, or a nitrogen protecting group, or two R^(N)groups are joined to form an optionally substituted heterocyclic ring,or one R^(N) group and one R^(D) group are joined to form an optionallysubstituted heterocyclic ring; each instance of R^(F) is independentlyhydrogen or halo; and each instance of R^(D) is independently hydrogen,halo, —CN, —NO₂, optionally substituted alkyl, optionally substitutedalkenyl, optionally substituted alkynyl, optionally substitutedcarbocyclyl, optionally substituted aryl, optionally substitutedheterocyclyl, optionally substituted heteroaryl, —OR^(A), —N(R^(B))₂,—SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A), —C(O)N(R^(B))₂,—C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂, —NR^(B)C(O)R^(A),—NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂, —NR^(B)C(O)OR^(A),—SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A),—C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B), —C(═S)R^(A),—C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A), —OS(O)₂R^(A), —SO₂R^(A),—NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or two R^(D) groups are joined toform an optionally substituted carbocyclic or optionally substitutedheterocyclic ring.
 80. The compound of claim 79, or a pharmaceuticallyacceptable salt thereof, wherein Formula (q-7)-(q-17) is of Formula(q-46)-(q-49):


81. The compound of claim 79, or a pharmaceutically acceptable saltthereof, wherein Formula (q-7)-(q-17) is of Formula (q-46a)-(q-49a):


82. The compound of claim 79, or a pharmaceutically acceptable saltthereof, wherein Formula (q-7)-(q-17) is of Formula (q-50a)-(q-57a):


83. The compound of claim 79, or a pharmaceutically acceptable saltthereof, wherein Formula (q-7)-(q-17) is of Formula (r-1) to (r-4):

wherein V²⁶, V²⁷, V²⁸, V²⁹, and V³⁰ are each independently O, S,NR^(Na), C═O, or C(R^(E))₂ as valency permits, provided no more than twoof V²⁶, V²⁷, V²⁸, V²⁹, and V³⁰ is O, S, or NR^(Na); or wherein one ofV²⁶, V²⁷, V²⁸, V²⁹, and V³⁰ and another of V²⁶, V²⁷, V²⁸, V²⁹, and V³⁰adjacent to each other are joined to form an N═C(R^(E)) orC(R^(E))═C(R^(E)) group; each instance of R^(E) is independentlyhydrogen, halo, —CN, —NO₂, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted aryl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or two R^(E)groups are joined to form an optionally substituted carbocyclic oroptionally substituted heterocyclic ring; and each instance of R^(Na) isindependently hydrogen, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, optionally substituted heteroaryl, —C(═O)R^(A),—C(═O)OR^(A), —C(═O)SR^(A), —C(═O)N(R^(B))₂, —C(═NR^(B))R^(A),—C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —C(═S)R^(A),—C(═S)N(R^(B))₂, —S(═O)R^(A), —SO₂R^(A), —SO₂N(R^(B))₂, or a nitrogenprotecting group
 84. The compound of claim 79, or a pharmaceuticallyacceptable salt thereof, wherein Formula (q-7)-(q-17) is of Formula(r-5) to (r-24):

wherein V²⁶, V²⁷, V²⁸, V²⁹, and V³⁰ are each independently O, S,NR^(Na), C═O, or C(R^(E))₂ as valency permits, provided no more than twoof V²⁶, V²⁷, V²⁸, V²⁹, and V³⁰ is O, S, or NR^(Na); or wherein one ofV²⁶, V²⁷, V²⁸, V²⁹, and V³⁰ and another of V²⁶, V²⁷, V²⁸, V²⁹, and V³⁰adjacent to each other are joined to form an N═C(R^(E)) orC(R^(E))═C(R^(E)) group; each instance of R^(E) is independentlyhydrogen, halo, —CN, —NO₂, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted aryl, optionallysubstituted heterocyclyl, optionally substituted heteroaryl, —OR^(A),—N(R^(B))₂, —SR^(A), —C(═O)R^(A), —C(O)OR^(A), —C(O)SR^(A),—C(O)N(R^(B))₂, —C(O)N(R^(B))N(R^(B))₂, —OC(O)R^(A), —OC(O)N(R^(B))₂,—NR^(B)C(O)R^(A), —NR^(B)C(O)N(R^(B))₂, —NR^(B)C(O)N(R^(B))N(R^(B))₂,—NR^(B)C(O)OR^(A), —SC(O)R^(A), —C(═NR^(B))R^(A), —C(═NNR^(B))R^(A),—C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —NR^(B)C(═NR^(B))R^(B),—C(═S)R^(A), —C(═S)N(R^(B))₂, —NR^(B)C(═S)R^(A), —S(O)R^(A),—OS(O)₂R^(A), —SO₂R^(A), —NR^(B)SO₂R^(A), or —SO₂N(R^(B))₂; or two R^(E)groups are joined to form an optionally substituted carbocyclic oroptionally substituted heterocyclic ring; and each instance of R^(Na) isindependently hydrogen, optionally substituted alkyl, optionallysubstituted alkenyl, optionally substituted alkynyl, optionallysubstituted carbocyclyl, optionally substituted heterocyclyl, optionallysubstituted aryl, optionally substituted heteroaryl, —C(═O)R^(A),—C(═O)OR^(A), —C(═O)SR^(A), —C(═O)N(R^(B))₂, —C(═NR^(B))R^(A),—C(═NNR^(B))R^(A), —C(═NOR^(A))R^(A), —C(═NR^(B))N(R^(B))₂, —C(═S)R^(A),—C(═S)N(R^(B))₂, —S(═O)R^(A), —SO₂R^(A), —SO₂N(R^(B))₂, or a nitrogenprotecting group.
 85. The compound of claim 84, or a pharmaceuticallyacceptable salt thereof, wherein Formula (r-5) to (r-24) is of Formula(r-148) to (r-161):